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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="author" content="Sarine Markossian, Abigail Grossman, Michelle Arkin, Douglas Auld, Chris Austin, Jonathan Baell, Kyle Brimacombe, Thomas D.Y. Chung, Nathan P. Coussens, Jayme L. Dahlin, Viswanath Devanarayan, Timothy L. Foley, Marcie Glicksman, Kirill Gorshkov, Joseph V. Haas, Matthew D. Hall, Samuel Hoare, James Inglese, Philip W. Iversen, Madhu Lal-Nag, Zhuyin Li, Jason R. Manro, James McGee, Owen McManus, Mackenzie Pearson, Terry Riss, Peter Saradjian, G. Sitta Sittampalam, Mike Tarselli, O. Joseph Trask, Jeffrey R. 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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/Book"><div class="meta-content fm-sec"><div class="iconblock whole_rhythm clearfix no_top_margin"><a href="http://www.ncats.nih.gov/" title="Eli Lilly & Company and the National Center for Advancing Translational Sciences" class="img_link icnblk_img" ref="pagearea=logo&targetsite=external&targetcat=link&targettype=publisher"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-assayguide-lrg.png" alt="Cover of Assay Guidance Manual" /></a><div class="icnblk_cntnt"><h1 id="_NBK53196_"><span itemprop="name">Assay Guidance Manual</span></h1><p class="contrib-group">Editors: <span itemprop="editor">Sarine Markossian</span>, Editor-in-Chief and <span itemprop="editor">Abigail Grossman</span>, Managing Editor. Associate Editors: <span itemprop="editor">Michelle Arkin</span>, <span itemprop="editor">Douglas Auld</span>, <span itemprop="editor">Chris Austin</span>, <span itemprop="editor">Jonathan Baell</span>, <span itemprop="editor">Kyle Brimacombe</span>, <span itemprop="editor">Thomas D.Y. Chung</span>, <span itemprop="editor">Nathan P. Coussens</span>, <span itemprop="editor">Jayme L. Dahlin</span>, <span itemprop="editor">Viswanath Devanarayan</span>, <span itemprop="editor">Timothy L. Foley</span>, <span itemprop="editor">Marcie Glicksman</span>, <span itemprop="editor">Kirill Gorshkov</span>, <span itemprop="editor">Joseph V. Haas</span>, <span itemprop="editor">Matthew D. Hall</span>, <span itemprop="editor">Samuel Hoare</span>, <span itemprop="editor">James Inglese</span>, <span itemprop="editor">Philip W. Iversen</span>, <span itemprop="editor">Madhu Lal-Nag</span>, <span itemprop="editor">Zhuyin Li</span>, <span itemprop="editor">Jason R. Manro</span>, <span itemprop="editor">James McGee</span>, <span itemprop="editor">Owen McManus</span>, <span itemprop="editor">Mackenzie Pearson</span>, <span itemprop="editor">Terry Riss</span>, <span itemprop="editor">Peter Saradjian</span>, <span itemprop="editor">G. Sitta Sittampalam</span>, <span itemprop="editor">Mike Tarselli</span>, <span itemprop="editor">O. Joseph Trask, Jr.</span>, <span itemprop="editor">Jeffrey R. Weidner</span>, <span itemprop="editor">Mary Jo Wildey</span>, <span itemprop="editor">Kelli Wilson</span>, <span itemprop="editor">Menghang Xia</span>, and <span itemprop="editor">Xin Xu</span>.</p><a data-jig="ncbitoggler" href="#__NBK53196_ai__" style="border:0;text-decoration:none">Editor Information</a><div style="display:none" class="ui-widget" id="__NBK53196_ai__"><p class="contrib-group"><h4>Editors</h4>Editors: <span itemprop="editor">Sarine Markossian</span>, Editor-in-Chief<sup>1</sup> and <span itemprop="editor">Abigail Grossman</span>, Managing Editor<sup>1</sup>. Associate Editors: <span itemprop="editor">Michelle Arkin</span>,<sup>2</sup> <span itemprop="editor">Douglas Auld</span>,<sup>3</sup> <span itemprop="editor">Chris Austin</span>,<sup>4</sup> <span itemprop="editor">Jonathan Baell</span>,<sup>5</sup> <span itemprop="editor">Kyle Brimacombe</span>,<sup>1</sup> <span itemprop="editor">Thomas D.Y. Chung</span>,<sup>6</sup> <span itemprop="editor">Nathan P. Coussens</span>,<sup>7</sup> <span itemprop="editor">Jayme L. Dahlin</span>,<sup>23</sup> <span itemprop="editor">Viswanath Devanarayan</span>,<sup>8</sup> <span itemprop="editor">Timothy L. Foley</span>,<sup>9</sup> <span itemprop="editor">Marcie Glicksman</span>,<sup>10</sup> <span itemprop="editor">Kirill Gorshkov</span>,<sup>11</sup> <span itemprop="editor">Joseph V. Haas</span>,<sup>12</sup> <span itemprop="editor">Matthew D. Hall</span>,<sup>1</sup> <span itemprop="editor">Samuel Hoare</span>,<sup>13</sup> <span itemprop="editor">James Inglese</span>,<sup>1</sup> <span itemprop="editor">Philip W. Iversen</span>,<sup>14</sup> <span itemprop="editor">Madhu Lal-Nag</span>,<sup>15</sup> <span itemprop="editor">Zhuyin Li</span>,<sup>11</sup> <span itemprop="editor">Jason R. Manro</span>,<sup>12</sup> <span itemprop="editor">James McGee</span>,<sup>12</sup> <span itemprop="editor">Owen McManus</span>,<sup>16</sup> <span itemprop="editor">Mackenzie Pearson</span>,<sup>12</sup> <span itemprop="editor">Terry Riss</span>,<sup>17</sup> <span itemprop="editor">Peter Saradjian</span>,<sup>18</sup> <span itemprop="editor">G. Sitta Sittampalam</span>,<sup>1</sup> <span itemprop="editor">Mike Tarselli</span>,<sup>19</sup> <span itemprop="editor">O. Joseph Trask, Jr.</span>,<sup>24</sup> <span itemprop="editor">Jeffrey R. Weidner</span>,<sup>21</sup> <span itemprop="editor">Mary Jo Wildey</span>,<sup>22</sup> <span itemprop="editor">Kelli Wilson</span>,<sup>1</sup> <span itemprop="editor">Menghang Xia</span>,<sup>1</sup> and <span itemprop="editor">Xin Xu</span><sup>1</sup>.</p><h4>Affiliations</h4><div class="affiliation"><sup>1</sup> National Center for Advancing Translational Sciences, National
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Institutes of Health</div><div class="affiliation"><sup>2</sup> University of California, San Francisco</div><div class="affiliation"><sup>3</sup> Novartis Institutes for Biomedical Research</div><div class="affiliation"><sup>4</sup> GSK plc</div><div class="affiliation"><sup>5</sup> Lyterian Therapeutics</div><div class="affiliation"><sup>6</sup> Sanford Burnham Prebys
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Medical Discovery Institute</div><div class="affiliation"><sup>7</sup> Frederick National Laboratory for Cancer Research</div><div class="affiliation"><sup>8</sup> Eisai, Inc</div><div class="affiliation"><sup>9</sup> Pfizer Inc</div><div class="affiliation"><sup>10</sup> EnClear Therapies</div><div class="affiliation"><sup>11</sup> Bristol Myers Squibb</div><div class="affiliation"><sup>12</sup> Eli Lilly and Company</div><div class="affiliation"><sup>13</sup> Pharmechanics, LLC</div><div class="affiliation"><sup>14</sup> Luther College</div><div class="affiliation"><sup>15</sup> InSphero AG</div><div class="affiliation"><sup>16</sup> Quiver Bioscience</div><div class="affiliation"><sup>17</sup> Promega Corporation</div><div class="affiliation"><sup>18</sup> Beth Israel Deaconess Medical Center</div><div class="affiliation"><sup>19</sup> TetraScience</div><div class="affiliation"><sup>20</sup> PerkinElmer, Inc.</div><div class="affiliation"><sup>21</sup> QualSci Consulting, LLC</div><div class="affiliation"><sup>22</sup> Merck & Co., Inc.</div><div class="affiliation"><sup>23</sup> Agios Pharmaceuticals</div><div class="affiliation"><sup>24</sup> Revvity, Inc.</div></div><div class="half_rhythm">Bethesda (MD): <a href="http://www.ncats.nih.gov/" ref="pagearea=meta&targetsite=external&targetcat=link&targettype=publisher"><span itemprop="publisher">Eli Lilly & Company and the National Center for Advancing
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Translational Sciences</span></a>; <span itemprop="datePublished">2004</span>-.</div><div class="half_rhythm"><ul class="inline_list"><li><span class="label"><a data-jig="ncbidialog" href="#_ncbi_dlg_cpyrght_NBK53196" data-jigconfig="modal:true">Copyright and
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Permissions</a></span></li></ul></div><div id="_ncbi_dlg_cpyrght_NBK53196" style="display:none" title="Copyright and Permissions"><div><div class="half_rhythm"><div>All Assay Guidance Manual content, except where otherwise noted, is
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licensed under a <a href="http://creativecommons.org/licenses/by-nc-sa/3.0/" ref="pagearea=meta&targetsite=external&targetcat=link&targettype=uri">Creative
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Commons Attribution-NonCommercial-ShareAlike 3.0 Unported</a> license
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(CC BY-NC-SA 3.0), which permits copying, distribution, transmission, and
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adaptation of the work, provided the original work is properly cited and not
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used for commercial purposes. Any altered, transformed, or adapted form of the
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work may only be distributed under the same or similar license to this
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one.</div></div></div></div><div class="bk_noprnt"><form method="get" action="/books/n/assayguide/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div></div></div><div class="body-content whole_rhythm" itemprop="text"><div itemprop="description"><p>This eBook is a comprehensive, crucial resource for investigators optimizing assays
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to evaluate collections of molecules with the overall goal of developing probes that
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modulate the activity of biological targets, pathways or cellular phenotypes. Such
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probes might be candidates for further optimization and investigation in drug
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discovery and development.</p><p>Originally written as a guide for therapeutic project teams within a major
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pharmaceutical company, this manual has been adapted to provide guidelines for
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scientists in academic, non-profit, government and industrial research laboratories
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to develop assay formats compatible with <a href="https://www.ncbi.nlm.nih.gov/books/n/assayguide/qbiogloss/def-item/high-throughput-screening-hts-/" ref="pagearea=abstract&targetsite=external&targetcat=link&targettype=uri">High Throughput Screening (HTS)</a> and Structure Activity Relationship
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(SAR) measurements of new and known molecular entities. Topics addressed in this
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manual include: <ul><li class="half_rhythm"><div>Descriptions of assay formats that are compatible with <a href="https://www.ncbi.nlm.nih.gov/books/n/assayguide/qbiogloss/def-item/high-throughput-screening-hts-/" ref="pagearea=abstract&targetsite=external&targetcat=link&targettype=uri">HTS</a> and determination of SAR</div></li><li class="half_rhythm"><div>Selection and development of optimal assay reagents</div></li><li class="half_rhythm"><div>Optimizations and troubleshooting for assay protocols with respect to
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sensitivity, dynamic range, signal intensity and stability</div></li><li class="half_rhythm"><div>Adaptations of assays for automation and scaling to microtiter plate
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formats</div></li><li class="half_rhythm"><div>Instrumentation </div></li><li class="half_rhythm"><div>Sources of assay artifacts and interferences </div></li><li class="half_rhythm"><div>Statistical validation of assay performance parameters</div></li><li class="half_rhythm"><div>Secondary assays for chemical probe validation and SAR refinement</div></li><li class="half_rhythm"><div>Data standards for reporting the results of screening and SAR assays</div></li><li class="half_rhythm"><div><i>In vivo</i> assay development and validation</div></li><li class="half_rhythm"><div>Assay development and validation for siRNA-based high-throughput
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screens</div></li></ul>
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</p><p>The <a href="https://ncats.nih.gov/" ref="pagearea=abstract&targetsite=external&targetcat=link&targettype=uri">National Center
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for Advancing Translational Sciences (NCATS)</a> manages the content of
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the Assay Guidance Manual with input from industry, academia and government experts.
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More than 100 authors from around the globe have contributed content to this free
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resource, which is updated quarterly with contributions by experienced scientists
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from multiple disciplines working in drug discovery and development worldwide.</p><p>For more information about the Assay Guidance Manual and related training
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opportunities, visit <a href="https://ncats.nih.gov/expertise/preclinical/agm" ref="pagearea=abstract&targetsite=external&targetcat=link&targettype=uri">https://ncats.nih.gov/expertise/preclinical/agm</a>.</p></div><div><h2>Contents</h2><div class="bktoc_all_cntnr top align_right" style="display:none"><ul class="inline_list_right"><li><a class="bktoc_all_exp" href="#">Expand All</a></li><li style="margin-left:.8em"><a class="bktoc_all_clps" href="#">Collapse All</a></li></ul></div><ul id="toc_tllNBK53196_toc_updates" class="simple-list toc toc-toggle"><li class="half_rhythm" id="toc_itm_NBK53196_toc_updates"><a class="toc-item" href="/books/n/assayguide/updates/">New in Assay Guidance Manual</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_preface"><a class="toc-item" href="/books/n/assayguide/preface/">Preface</a><div>G. Sitta Sittampalam and Nathan P. Coussens.</div><div class="small">Published May 1, 2012; Last Update: March 31, 2017.</div></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_acknowledgements"><a class="toc-item" href="/books/n/assayguide/acknowledgements/">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_intro_xml"><a class="toc-item" href="/books/n/assayguide/intro.xml/">Considerations for Early Phase Drug Discovery</a><div>Editors: Thomas D.Y. Chung, Nathan P. Coussens, G. Sitta Sittampalam, and Yueming Wang.</div><ul id="toc_lst_NBK53196_toc_intro_xml" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/">Early Drug Discovery and Development Guidelines: For Academic Researchers, Collaborators, and Start-up Companies</a><div>Jeffrey Strovel, Sitta Sittampalam, Nathan P. Coussens, Michael Hughes, James Inglese, Andrew Kurtz, Ali Andalibi, Lavonne Patton, Chris Austin, Michael Baltezor, Michael Beckloff, Michael Weingarten, and Scott Weir.</div><div class="small">Published May 1, 2012; Last Update: July 1, 2016.</div><ul id="toc_lst_NBK53196_toc_drugdiscovery" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Abstract"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Background"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Background">Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Purpose"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Purpose">Purpose</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Scope"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Scope">Scope</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Assumptions"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Assumptions">Assumptions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Definitions"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Definitions">Definitions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Discovery_and_Development"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Discovery_and_Development">Section 1. Discovery and Development of New Chemical Entities</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Repurposing_of_Marketed_Dr"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Repurposing_of_Marketed_Dr">Section 2. Repurposing of Marketed Drugs</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Development_of_Drug_Delive"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Development_of_Drug_Delive">Section 3. Development of Drug Delivery Platform Technology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Alternative_NCE_Strategy_E"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Alternative_NCE_Strategy_E">Section 4. Alternative NCE Strategy: Exploratory IND</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Orphan_Drug_Designation"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Orphan_Drug_Designation">Section 5. Orphan Drug Designation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_Conclusion"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_drugdiscovery_References"><a class="toc-item" href="/books/n/assayguide/drugdiscovery/#drugdiscovery.References">References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitroba"><a class="toc-item" href="/books/n/assayguide/invitroba/">
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<i>In Vitro</i> Biochemical Assays</a><div>Editors: Nathan P. Coussens, Timothy L. Foley, Sam R.J. Hoare, James McGee, and G. Sitta Sittampalam.</div><ul id="toc_lst_NBK53196_toc_invitroba" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding"><a class="toc-item" href="/books/n/assayguide/kineticbinding/">Analyzing Kinetic Binding Data</a><div>Sam R.J. Hoare.</div><div class="small">Published April 1, 2021.</div><ul id="toc_lst_NBK53196_toc_kineticbinding" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Abstract"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Introduction"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Basic_Principles_of_Ligan"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Basic_Principles_of_Ligan">Basic Principles of Ligand Binding Kinetics</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Kinetic_Analysis_Methods"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Kinetic_Analysis_Methods">Kinetic Analysis Methods for Direct Target-Ligand Binding Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Competition_Kinetics_Quan"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Competition_Kinetics_Quan">Competition Kinetics: Quantifying Kinetics by Competition Against a Tracer Ligand</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Compound_Washout_Method"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Compound_Washout_Method">Compound Washout Method</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Complex_Binding_Mechanism"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Complex_Binding_Mechanism">Complex Binding Mechanisms</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Binding_Kinetics_from_Fun"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Binding_Kinetics_from_Fun">Binding Kinetics from Functional Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Summary"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Summary">Summary</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_Appendix_Equations_for_Bi"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.Appendix_Equations_for_Bi">Appendix: Equations for Binding Kinetic Data Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_kineticbinding_References"><a class="toc-item" href="/books/n/assayguide/kineticbinding/#kineticbinding.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp"><a class="toc-item" href="/books/n/assayguide/vimp/">Validating Identity, Mass Purity and Enzymatic Purity of Enzyme Preparations</a><div>John E Scott and Kevin P Williams.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_vimp" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Abstract"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Definitions"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Definitions">Definitions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Consequences_of_using_Enzymatically"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Consequences_of_using_Enzymatically">Consequences of using Enzymatically Impure Enzyme Preparations.</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Signs_of_Enzymatic_Contamination"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Signs_of_Enzymatic_Contamination">Signs of Enzymatic Contamination</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Solutions_for_Enzymatic_Contaminati"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Solutions_for_Enzymatic_Contaminati">Solutions for Enzymatic Contamination</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Importance_of_Batch_Testing"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Importance_of_Batch_Testing">Importance of Batch Testing</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Identity_and_Mass_Purity"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Identity_and_Mass_Purity">Identity and Mass Purity</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Assay_Design_Factors_that_Affect_th"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Assay_Design_Factors_that_Affect_th">Assay Design Factors that Affect the Likelihood of Detecting Enzyme Impurities</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_Validating_Enzymatic_Purity"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.Validating_Enzymatic_Purity">Validating Enzymatic Purity</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_vimp_References"><a class="toc-item" href="/books/n/assayguide/vimp/#vimp.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics"><a class="toc-item" href="/books/n/assayguide/eabasics/">Basics of Enzymatic Assays for HTS</a><div>Harold B. Brooks, Sandaruwan Geeganage, Steven D. Kahl, Chahrzad Montrose, Sitta Sittampalam, Michelle C. Smith, and Jeffrey R. Weidner.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_eabasics" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Abstract"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Enzyme_Assay_Development_Flow_C"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Enzyme_Assay_Development_Flow_C">Enzyme Assay Development Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Introduction"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Concept"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Concept">Concept</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Reagents_and_Method_Development"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Reagents_and_Method_Development">Reagents and Method Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Detection_System_Linearity"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Detection_System_Linearity">Detection System Linearity</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Enzyme_Reaction_Progress_Curve"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Enzyme_Reaction_Progress_Curve">Enzyme Reaction Progress Curve</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Measurement_of_Km_and_Vmax"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Measurement_of_Km_and_Vmax">Measurement of K<sub>m</sub> and V<sub>max</sub></a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Determination_of_IC50_for_Inhib"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Determination_of_IC50_for_Inhib">Determination of IC<sub>50</sub> for Inhibitors</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_IC50_Determination_for_SAR"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.IC50_Determination_for_SAR">IC<sub>50</sub> Determination for SAR</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Optimization_Experiments"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Optimization_Experiments">Optimization Experiments</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_Assay_Validation"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.Assay_Validation">Assay Validation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_eabasics_References"><a class="toc-item" href="/books/n/assayguide/eabasics/#eabasics.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa"><a class="toc-item" href="/books/n/assayguide/enzymemoa/">Mechanism of Action Assays for Enzymes</a><div>John Strelow, Walthere Dewe, Phillip W Iversen, Harold B Brooks, Jeffrey A Radding, James McGee, and Jeffrey Weidner.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_enzymemoa" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_Abstract"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_Overview_of_MOA_in_Drug_Discov"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.Overview_of_MOA_in_Drug_Discov">Overview of MOA in Drug Discovery</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_Types_of_Inhibition"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.Types_of_Inhibition">Types of Inhibition</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_Performing_MOA_Studies"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.Performing_MOA_Studies">Performing MOA Studies</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_References"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_enzymemoa_Glossary_of_MOA_Terms"><a class="toc-item" href="/books/n/assayguide/enzymemoa/#enzymemoa.Glossary_of_MOA_Terms">Glossary of MOA Terms</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke"><a class="toc-item" href="/books/n/assayguide/assaydevpke/">Assay Development for Protein Kinase Enzymes</a><div>J. Fraser Glickman.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_assaydevpke" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Abstract"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Introduction"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Assay_Development"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Assay_Development">Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Considerations_of_Mechanism"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Considerations_of_Mechanism">Considerations of Mechanism</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Assay_Optimization"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Assay_Optimization">Assay Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_References"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaydevpke_Additional_References"><a class="toc-item" href="/books/n/assayguide/assaydevpke/#assaydevpke.Additional_References">Additional References:</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding"><a class="toc-item" href="/books/n/assayguide/receptorbinding/">Receptor Binding Assays for HTS and Drug Discovery</a><div>Douglas S Auld, Mark W. Farmen, Steven D. Kahl, Aidas Kriauciunas, Kevin L. McKnight, Chahrzad Montrose, and Jeffrey R. Weidner.</div><div class="small">Published May 1, 2012; Last Update: July 1, 2018.</div><ul id="toc_lst_NBK53196_toc_receptorbinding" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Abstract"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Introduction"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Flow_Chart_of_Steps_to_A"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Flow_Chart_of_Steps_to_A">Flow Chart of Steps to Assay Development for SPA Format</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Flow_Chart_of_Steps_to_A_1"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Flow_Chart_of_Steps_to_A_1">Flow Chart of Steps to Assay Development for Filter Format</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Scintillation_Proximity"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Scintillation_Proximity">Scintillation Proximity Assays (SPA)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_SPA_Assay_Format"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.SPA_Assay_Format">SPA Assay Format</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Assay_Buffer"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Assay_Buffer">Assay Buffer</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Solvent_Interference_Con"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Solvent_Interference_Con">Solvent Interference Conditions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Binding_Parameter"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Binding_Parameter">Binding Parameter</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Filtration_Assays"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Filtration_Assays">Filtration Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Filter_Assay_Format"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Filter_Assay_Format">Filter Assay Format</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Assay_Buffer_Filter"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Assay_Buffer_Filter">Assay Buffer (Filter)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Assay_Conditions_Filter"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Assay_Conditions_Filter">Assay Conditions (Filter)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Binding_Parameters_Filte"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Binding_Parameters_Filte">Binding Parameters (Filter)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_Practical_Use_of_Fluores"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.Practical_Use_of_Fluores">Practical Use of Fluorescence Polarization in Competitive Receptor Binding Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_receptorbinding_References"><a class="toc-item" href="/books/n/assayguide/receptorbinding/#receptorbinding.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev"><a class="toc-item" href="/books/n/assayguide/proteaserev/">Protease Assays</a><div>Guofeng Zhang.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_proteaserev" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Abstract"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Introduction"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Homogenous_Assays"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Homogenous_Assays">Homogenous Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_SeparationBased_Assays"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.SeparationBased_Assays">Separation-Based Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Triaging_Assay_Hits"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Triaging_Assay_Hits">Triaging Assay Hits</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Data_Analysis_for_Time_Depen"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Data_Analysis_for_Time_Depen">Data Analysis for Time Dependent Inhibitors and Covalent Inhibitors</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_Conclusion"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_proteaserev_References"><a class="toc-item" href="/books/n/assayguide/proteaserev/#proteaserev.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi"><a class="toc-item" href="/books/n/assayguide/ppi/">Inhibition of Protein-Protein Interactions: Non-Cellular Assay Formats</a><div>Michelle R. Arkin, Marcie A. Glicksman, Haian Fu, Jonathan J. Havel, and Yuhong Du.</div><div class="small">Published March 18, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_ppi" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Abstract"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Overview_and_Introduction"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Overview_and_Introduction">Overview and Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_ELISAtype_assays"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.ELISAtype_assays">ELISA-type assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Mixandread_assays"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Mixandread_assays">Mix-and-read assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Validating_druglike_binding_of_PPI_i"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Validating_druglike_binding_of_PPI_i">Validating drug-like binding of PPI inhibitors</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Useful_websites"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Useful_websites">Useful websites</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_References"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_Additional_References"><a class="toc-item" href="/books/n/assayguide/ppi/#ppi.Additional_References">Additional References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh"><a class="toc-item" href="/books/n/assayguide/immunometh/">Immunoassay Methods</a><div>Karen L. Cox, Viswanath Devanarayan, Aidas Kriauciunas, Joseph Manetta, Chahrzad Montrose, and Sitta Sittampalam.</div><div class="small">Published May 1, 2012; Last Update: July 8, 2019.</div><ul id="toc_lst_NBK53196_toc_immunometh" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Abstract"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Immunoassay_Development_Optim"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Immunoassay_Development_Optim">Immunoassay Development, Optimization and Validation Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Introduction"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Immunoassay_Parameters"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Immunoassay_Parameters">Immunoassay Parameters</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Reagents"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Reagents">Reagents</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Instrumentation"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Instrumentation">Instrumentation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Immunoassay_Formats"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Immunoassay_Formats">Immunoassay Formats</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Important_Parameters_for_Deve"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Important_Parameters_for_Deve">Important Parameters for Development of an Immunoassay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Initial_Concept_and_Method_De"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Initial_Concept_and_Method_De">Initial Concept and Method Development for a Sandwich Immunoassay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Initial_Concept_and_Method_De_1"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Initial_Concept_and_Method_De_1">Initial Concept and Method Development of a Competitive Assay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Development_of_a_Competitive"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Development_of_a_Competitive">Development of a Competitive Immunoassay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Method_Validation_PreStudy"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Method_Validation_PreStudy">Method Validation (Pre-Study)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Method_Validation_InStudy"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Method_Validation_InStudy">Method Validation (In-Study)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_PreStudy__InStudy_Acceptance"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.PreStudy__InStudy_Acceptance">Pre-Study & In-Study Acceptance Criteria</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Precision_Profile_Tool_for_Es"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Precision_Profile_Tool_for_Es">Precision Profile Tool for Estimating Assay Performance Characteristics</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Immunogenicity_Cut_Point_Anal"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Immunogenicity_Cut_Point_Anal">Immunogenicity Cut Point Analysis Tool (I-CAT)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_References"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_immunometh_Additional_References"><a class="toc-item" href="/books/n/assayguide/immunometh/#immunometh.Additional_References">Additional References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp"><a class="toc-item" href="/books/n/assayguide/gtp/">GTPγS Binding Assays</a><div>Neil W. DeLapp, Wendy H. Gough, Steven D. Kahl, Amy C. Porter, and Todd R. Wiernicki.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_gtp" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Abstract"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Flow_Chart_for_Assay_Development"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Flow_Chart_for_Assay_Development">Flow Chart for Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Introduction"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Materials_and_Reagents"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Materials_and_Reagents">Materials and Reagents</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Membrane_Preparations_and_Assay_Buff"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Membrane_Preparations_and_Assay_Buff">Membrane Preparations and Assay Buffers</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Basic_Assay_Protocol"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Basic_Assay_Protocol">Basic Assay Protocol</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Assay_Optimization"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Assay_Optimization">Assay Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Data_Analysis"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Data_Analysis">Data Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Filtration_Assays"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Filtration_Assays">Filtration Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_NonRadiometric_GTP_S_Assays"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.NonRadiometric_GTP_S_Assays">Non-Radiometric GTPγS Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_References"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gtp_Additional_References"><a class="toc-item" href="/books/n/assayguide/gtp/#gtp.Additional_References">Additional References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat"><a class="toc-item" href="/books/n/assayguide/hat/">Histone Acetyltransferase Assays in Drug and Chemical Probe Discovery</a><div>Sanket Gadhia, Jonathan H. Shrimp, Jordan L. Meier, James E. McGee, and Jayme L. Dahlin.</div><div class="small">Published July 26, 2017; Last Update: July 1, 2018.</div><ul id="toc_lst_NBK53196_toc_hat" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Abstract"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Flowchart"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Abbreviations"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Introduction_and_Background"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Introduction_and_Background">Introduction and Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_General_Considerations_for_HAT_Assay"><a class="toc-item" href="/books/n/assayguide/hat/#hat.General_Considerations_for_HAT_Assay">General Considerations for HAT Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Biochemical_HAT_Assays"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Biochemical_HAT_Assays">Biochemical HAT Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Section_Summary_1"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Section_Summary_1">Section Summary</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_CellBased_HAT_Assays"><a class="toc-item" href="/books/n/assayguide/hat/#hat.CellBased_HAT_Assays">Cell-Based HAT Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Miscellaneous_Considerations"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Miscellaneous_Considerations">Miscellaneous Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Conclusions"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Conflict_of_Interest_Statement"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Conflict_of_Interest_Statement">Conflict of Interest Statement</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_Suggesting_Readings_alphabetical_ord"><a class="toc-item" href="/books/n/assayguide/hat/#hat.Suggesting_Readings_alphabetical_ord">Suggesting Readings (alphabetical order)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hat_References"><a class="toc-item" href="/books/n/assayguide/hat/#hat.References">References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivcba_xml"><a class="toc-item" href="/books/n/assayguide/ivcba.xml/">
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<i>In Vitro</i> Cell Based Assays </a><div>Editors: Marcie Glicksman, Zhuyin Li, Sarine Markossian, Terry Riss, G. Sitta Sittampalam, O. Joseph Trask, Jr., and Yueming Wang.</div><ul id="toc_lst_NBK53196_toc_ivcba_xml" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml"><a class="toc-item" href="/books/n/assayguide/ml/">Machine Learning and Assay Development for Image-based Phenotypic Profiling of Drug Treatments</a><div>Jonathan Z. Sexton, Reid Fursmidt, Matthew J. O’Meara, Wienand Omta, Arvind Rao, David A. Egan, and Steven A. Haney.</div><div class="small">Published March 15, 2023.</div><ul id="toc_lst_NBK53196_toc_ml" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Abstract"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Flow_Chart"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Flow_Chart">Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Introduction"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Methods_and_Resources"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Methods_and_Resources">Methods and Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Protocol_Overview"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Protocol_Overview">Protocol Overview</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Image_Analysis_Getting_from_Images_to"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Image_Analysis_Getting_from_Images_to">Image Analysis: Getting from Images to Numbers</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Exploratory_Data_Analysis_in_Pilot_Sc"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Exploratory_Data_Analysis_in_Pilot_Sc">Exploratory Data Analysis in Pilot Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Machine_Learning_for_a_Group_of_Plate"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Machine_Learning_for_a_Group_of_Plate">Machine Learning for a Group of Plates</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Machine_Learning_Incorporating_Novel"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Machine_Learning_Incorporating_Novel">Machine Learning, Incorporating Novel Phenotype Identification, With Applications to Cell Painting and Related Phenotypic Screens</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_Conclusions"><a class="toc-item" href="/books/n/assayguide/ml/#ml.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ml_References"><a class="toc-item" href="/books/n/assayguide/ml/#ml.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays"><a class="toc-item" href="/books/n/assayguide/biosensorassays/">Biosensor Assays for Measuring the Kinetics of G-Protein and Arrestin-Mediated Signaling in Live Cells</a><div>Sam R.J. Hoare and Thomas E. Hughes.</div><div class="small">Published September 1, 2021.</div><ul id="toc_lst_NBK53196_toc_biosensorassays" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Abstract"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Introduction"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Measuring_GPCR_Signaling"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Measuring_GPCR_Signaling">Measuring GPCR Signaling Using Genetically-Encoded Fluorescent Biosensors</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Analyzing_Signaling_Time"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Analyzing_Signaling_Time">Analyzing Signaling Time Course Data</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Simplifying_Biased_Agoni"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Simplifying_Biased_Agoni">Simplifying Biased Agonism Assessment Using Signaling Kinetics</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Conclusions"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_biosensorassays_References"><a class="toc-item" href="/books/n/assayguide/biosensorassays/#biosensorassays.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_mttassays"><a class="toc-item" href="/books/n/assayguide/mttassays/">Cell Viability Assays</a><div>Terry L Riss, Richard A Moravec, Andrew L Niles, Sarah Duellman, Hélène A Benink, Tracy J Worzella, and Lisa Minor.</div><div class="small">Published May 1, 2013; Last Update: July 1, 2016.</div><ul id="toc_lst_NBK53196_toc_mttassays" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_mttassays_Abstract"><a class="toc-item" href="/books/n/assayguide/mttassays/#mttassays.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_mttassays_Introduction"><a class="toc-item" href="/books/n/assayguide/mttassays/#mttassays.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_mttassays_Conclusion"><a class="toc-item" href="/books/n/assayguide/mttassays/#mttassays.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_mttassays_References"><a class="toc-item" href="/books/n/assayguide/mttassays/#mttassays.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox"><a class="toc-item" href="/books/n/assayguide/cytotox/">Cytotoxicity Assays: <i>In Vitro</i> Methods to Measure Dead Cells</a><div>Terry Riss, Andrew Niles, Rich Moravec, Natashia Karassina, and Jolanta Vidugiriene.</div><div class="small">Published May 1, 2019.</div><ul id="toc_lst_NBK53196_toc_cytotox" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Abstract"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Introduction"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Dyes_That_Selectively_Penetrate"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Dyes_That_Selectively_Penetrate">Dyes That Selectively Penetrate Dead Cells</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Fluorescent_DNA_Binding_Dyes_Tha"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Fluorescent_DNA_Binding_Dyes_Tha">Fluorescent DNA Binding Dyes That Penetrate Dead Cells</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Optional_Protocol_to_Estimate_To"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Optional_Protocol_to_Estimate_To">Optional Protocol to Estimate Total Number of Cells</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Markers_That_Leak_Out_of_the_Cyt"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Markers_That_Leak_Out_of_the_Cyt">Markers That Leak Out of the Cytoplasm of Dead Cells into Culture Medium</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_Conclusion"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cytotox_References"><a class="toc-item" href="/books/n/assayguide/cytotox/#cytotox.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis"><a class="toc-item" href="/books/n/assayguide/apoptosis/">Apoptosis Marker Assays for HTS</a><div>Terry L. Riss, Martha A. O’Brien, Richard A. Moravec, Kevin Kupcho, and Andrew L. Niles.</div><div class="small">Published July 1, 2021.</div><ul id="toc_lst_NBK53196_toc_apoptosis" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Abstract"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Abbreviations"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Introduction"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Caspase_Activity_Detection"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Caspase_Activity_Detection">Caspase Activity Detection</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Protocol_to_Detect_Caspase37_A"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Protocol_to_Detect_Caspase37_A">Protocol to Detect Caspase-3/7 Activity as a Marker of Apoptosis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Detection_of_PS_Exposure_as_a"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Detection_of_PS_Exposure_as_a">Detection of PS Exposure as a Marker of Apoptosis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Protocol_to_Detect_PS_Exposure"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Protocol_to_Detect_PS_Exposure">Protocol to Detect PS Exposure as a Marker of Apoptosis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Data_Calculation"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Data_Calculation">Data Calculation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Commercial_Reagents"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Commercial_Reagents">Commercial Reagents</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_Conclusion"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_apoptosis_References"><a class="toc-item" href="/books/n/assayguide/apoptosis/#apoptosis.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell"><a class="toc-item" href="/books/n/assayguide/stemcell/">Pluripotent Stem Cell Culture Scale-Out</a><div>Debra L. Johnson, Robert Scott, and Rhonda Newman.</div><div class="small">Published June 1, 2021.</div><ul id="toc_lst_NBK53196_toc_stemcell" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Abstract"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Overview_and_Flowchart_of_Assay"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Overview_and_Flowchart_of_Assay">Overview and Flowchart of Assay Guidelines</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Introduction"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Stem_Cell_Culture_Consideration"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Stem_Cell_Culture_Consideration">Stem Cell Culture Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Assay_Protocol_Scaleout_of_hPSC"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Assay_Protocol_Scaleout_of_hPSC">Assay Protocol: Scale-out of hPSCs in Monolayer Culture Conditions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Monitoring_Cell_Health_in_Large"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Monitoring_Cell_Health_in_Large">Monitoring Cell Health in Large-Scale Culture</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_Conclusion"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_stemcell_References"><a class="toc-item" href="/books/n/assayguide/stemcell/#stemcell.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/">Authentication of Human and Mouse Cell Lines by Short Tandem Repeat (STR) DNA Genotype Analysis</a><div>Jamie L. Almeida and Christopher T. Korch.</div><div class="small">Published January 17, 2023.</div><ul id="toc_lst_NBK53196_toc_dnaprofiling" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Abstract"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Introduction"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Authentication_of_Mouse_Sam"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Authentication_of_Mouse_Sam">Authentication of Mouse Samples by STR Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_STR_DNA_genotyping_assays_f"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.STR_DNA_genotyping_assays_f">STR DNA genotyping assays for authentication of human and mouse cell line identities and detection of cross-contaminating intraspecies cells</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Data_Analysis_and_Interpret"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Data_Analysis_and_Interpret">Data Analysis and Interpretation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Criteria_for_Determining_Qu"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Criteria_for_Determining_Qu">Criteria for Determining Quality STR Profile Analysis for Reliable and Interpretable Results</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Services_for_STR_Genotyping"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Services_for_STR_Genotyping">Services for STR Genotyping of Cell Lines</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Troubleshooting"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Troubleshooting">Troubleshooting</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Noncommercial_Kits_and_Inap"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Noncommercial_Kits_and_Inap">Non-commercial Kits and Inappropriate Authentication Methods</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Glossary_of_Terms"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Glossary_of_Terms">Glossary of Terms</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_License"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.License">License</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_Useful_Resources"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.Useful_Resources">Useful Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dnaprofiling_References"><a class="toc-item" href="/books/n/assayguide/dnaprofiling/#dnaprofiling.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d"><a class="toc-item" href="/books/n/assayguide/invitro3d/">
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<i>In vitro</i> 3D Spheroids and Microtissues: ATP-based Cell Viability and Toxicity Assays</a><div>Monika Kijanska and Jens Kelm.</div><div class="small">Published January 21, 2016.</div><ul id="toc_lst_NBK53196_toc_invitro3d" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_Abstract"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_Flow_Chart_3D_Spheroid_and_Mic"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.Flow_Chart_3D_Spheroid_and_Mic">Flow Chart: 3D Spheroid and Microtissue Growth and Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_Introduction"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_sec1"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.sec1">1. <i>In Vitro</i> Toxicity and Drug Efficacy Testing in a 3D Spheroid Model</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_sec2"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.sec2">2. 3D Microtissue Viability Assay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invitro3d_References"><a class="toc-item" href="/books/n/assayguide/invitro3d/#invitro3d.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai"><a class="toc-item" href="/books/n/assayguide/cbrnai/">Cell-Based RNAi Assay Development for HTS</a><div>Scott Martin, Gene Buehler, Kok Long Ang, Farhana Feroze, Gopinath Ganji, and Yue Li.</div><div class="small">Published May 1, 2012; Last Update: May 1, 2013.</div><ul id="toc_lst_NBK53196_toc_cbrnai" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_Abstract"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_Introduction"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_OffTarget_Effects"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.OffTarget_Effects">Off-Target Effects</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_LossofFunction_Screens_Using_siRN"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.LossofFunction_Screens_Using_siRN">Loss-of-Function Screens Using siRNA</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_siRNA_Hit_Selection"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.siRNA_Hit_Selection">siRNA Hit Selection</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_RNAi_Synthetic_Lethality_Screens"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.RNAi_Synthetic_Lethality_Screens">RNAi Synthetic Lethality Screens</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_LossofFunction_Screens_Using_shRN"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.LossofFunction_Screens_Using_shRN">Loss-of-Function Screens Using <b>sh</b>RNA</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_Appendix"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.Appendix">Appendix</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_Abbreviations"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cbrnai_References"><a class="toc-item" href="/books/n/assayguide/cbrnai/#cbrnai.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr"><a class="toc-item" href="/books/n/assayguide/flipr/">FLIPR™ Assays for GPCR and Ion Channel Targets</a><div>Michelle R. Arkin, Patrick R. Connor, Renee Emkey, Kim E. Garbison, Beverly A. Heinz, Todd R. Wiernicki, Paul A. Johnston, Ramani A. Kandasamy, Nancy B. Rankl, and Sitta Sittampalam.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_flipr" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Abstract"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Overview_FLIPR_Assay_Development"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Overview_FLIPR_Assay_Development">Overview: FLIPR™ Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Introduction"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Types_of_FLIPR_Formats"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Types_of_FLIPR_Formats">Types of FLIPR™ Formats</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Reagents_and_Buffers_for_Method_De"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Reagents_and_Buffers_for_Method_De">Reagents and Buffers for Method Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Method_Development_and_Optimizatio"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Method_Development_and_Optimizatio">Method Development and Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_FLIPR_Instrument_Setup"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.FLIPR_Instrument_Setup">FLIPR Instrument Setup</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_Potential_Artifacts"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.Potential_Artifacts">Potential Artifacts</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_flipr_References"><a class="toc-item" href="/books/n/assayguide/flipr/#flipr.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel"><a class="toc-item" href="/books/n/assayguide/ionchannel/">Ion Channel Screening</a><div>Owen B McManus, Maria L Garcia, David Weaver, Melanie Bryant, Steven Titus, and James B Herrington.</div><div class="small">Published October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_ionchannel" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_Abstract"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_sec1"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.sec1">1. Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_sec2"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.sec2">2. Fluorescence Assays Using Membrane Potential Sensing Dyes</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_sec3"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.sec3">3. Ion Flux Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_sec4"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.sec4">4. HTS Assay Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_sec5"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.sec5">5. Preparation of Cells for Automated Electrophysiology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ionchannel_References"><a class="toc-item" href="/books/n/assayguide/ionchannel/#ionchannel.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea"><a class="toc-item" href="/books/n/assayguide/aea/">Automated Electrophysiology Assays</a><div>Birgit T. Priest, Rok Cerne, Michael J. Krambis, William A. Schmalhofer, Mark Wakulchik, Benjamin Wilenkin, and Kevin D. Burris.</div><div class="small">Published March 9, 2017.</div><ul id="toc_lst_NBK53196_toc_aea" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Introduction"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Flowchart"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Choice_of_Instrumentation"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Choice_of_Instrumentation">Choice of Instrumentation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Choice_of_Cell_Line"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Choice_of_Cell_Line">Choice of Cell Line</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Choice_of_Solutions"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Choice_of_Solutions">Choice of Solutions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_HighThroughput_Automated_Electrophys"><a class="toc-item" href="/books/n/assayguide/aea/#aea.HighThroughput_Automated_Electrophys">High-Throughput Automated Electrophysiology Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_MediumThroughput_Automated_Electroph"><a class="toc-item" href="/books/n/assayguide/aea/#aea.MediumThroughput_Automated_Electroph">Medium-Throughput Automated Electrophysiology Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_LowThroughput_Automated_Electrophysi"><a class="toc-item" href="/books/n/assayguide/aea/#aea.LowThroughput_Automated_Electrophysi">Low-Throughput Automated Electrophysiology Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Cost_of_Operation"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Cost_of_Operation">Cost of Operation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_References"><a class="toc-item" href="/books/n/assayguide/aea/#aea.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Appendix_1_Protocol_for_Assessment_o"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Appendix_1_Protocol_for_Assessment_o">Appendix 1: Protocol for Assessment of Use-Dependent Sodium Channel Inhibitors on IonWorks Quattro</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Appendix_2_Protocol_for_Assessment_o"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Appendix_2_Protocol_for_Assessment_o">Appendix 2: Protocol for Assessment of KCNQ1/KCNE1 Currents on IonWorks Barracuda</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Appendix_3_Protocol_for_Assessment_o"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Appendix_3_Protocol_for_Assessment_o">Appendix 3: Protocol for Assessment of GABA<sub>A</sub> Current Block on IonWorks Barracuda</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aea_Appendix_4_Protocol_for_Assessment_o"><a class="toc-item" href="/books/n/assayguide/aea/#aea.Appendix_4_Protocol_for_Assessment_o">Appendix 4: Protocol for Assessment of State-Dependent Sodium Channel Inhibitors on PatchXpress</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent"><a class="toc-item" href="/books/n/assayguide/highcontent/">Assay Development Guidelines for Image-Based High Content Screening, High Content Analysis and High Content Imaging</a><div>William Buchser, Mark Collins, Tina Garyantes, Rajarshi Guha, Steven Haney, Vance Lemmon, Zhuyin Li, and O. Joseph Trask.</div><div class="small">Published October 1, 2012; Last Update: September 22, 2014.</div><ul id="toc_lst_NBK53196_toc_highcontent" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Abstract"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Introduction"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Introduction">1. Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Image_Technologies_and_Instr"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Image_Technologies_and_Instr">2. Image Technologies and Instruments</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Assay_Concept_and_Design"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Assay_Concept_and_Design">3. Assay Concept and Design</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Cellular_Models_for_High_Con"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Cellular_Models_for_High_Con">4. Cellular Models for High Content Experiments</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Assay_Development_Considerat"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Assay_Development_Considerat">5. Assay Development Considerations and TroubleShooting</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Image_Acquisition_Analysis_a"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Image_Acquisition_Analysis_a">6. Image Acquisition, Analysis and Data Interpretation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Assay_Validation_for_HCA_Scr"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Assay_Validation_for_HCA_Scr">7. Assay Validation for HCA Screens</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_Data_Management_for_High_Con"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.Data_Management_for_High_Con">8. Data Management for High Content Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_highcontent_References"><a class="toc-item" href="/books/n/assayguide/highcontent/#highcontent.References">9. References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage"><a class="toc-item" href="/books/n/assayguide/hcsimage/">Advanced Assay Development Guidelines for Image-Based High Content Screening and Analysis</a><div>Mark-Anthony Bray, Anne Carpenter; Imaging Platform, Broad Institute of MIT and Harvard.</div><div class="small">Published July 8, 2017.</div><ul id="toc_lst_NBK53196_toc_hcsimage" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Abstract"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Experimental_design_for_HCS"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Experimental_design_for_HCS">1. Experimental design for HCS</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Assay_Quality_AND_Acceptance_Cr"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Assay_Quality_AND_Acceptance_Cr">2. Assay Quality AND Acceptance Criteria for HCS</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Quality_Control_for_HCS"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Quality_Control_for_HCS">3. Quality Control for HCS</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Normalization_of_HCS_data"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Normalization_of_HCS_data">4. Normalization of HCS data</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Measurement_of_image_features"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Measurement_of_image_features">5. Measurement of image features</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Machine_learning_for_HCS"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Machine_learning_for_HCS">6. Machine learning for HCS</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Wholeorganism_HCS"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Wholeorganism_HCS">7. Whole-organism HCS</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.Acknowledgements">8. Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsimage_References"><a class="toc-item" href="/books/n/assayguide/hcsimage/#hcsimage.References">9. References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb"><a class="toc-item" href="/books/n/assayguide/nfkb/">Nuclear Factor Kappa B (NF-κB) Translocation Assay Development and Validation for High Content Screening</a><div>O. Joseph Trask, Jr.</div><div class="small">Published October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_nfkb" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Abstract"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Introduction_and_background"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Introduction_and_background">Introduction and background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Assay_Development_Considerations"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Assay_Development_Considerations">Assay Development Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Assay_Development"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Assay_Development">Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Reference_Compounds"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Reference_Compounds">Reference Compounds</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Plate_Layout"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Plate_Layout">Plate Layout</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Protocol_for_Activators_Agonist_of"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Protocol_for_Activators_Agonist_of">Protocol for Activators (Agonist) of NFκB Pathway</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Logistics_Analysis_of_Protocol"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Logistics_Analysis_of_Protocol">Logistics Analysis of Protocol</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Cell_Based_Screen"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Cell_Based_Screen">Cell Based Screen</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Screening_Commercially_Available_NF"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Screening_Commercially_Available_NF">Screening Commercially Available NF-κB Antibodies (Appendix 1)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Safety_Considerations_Guidelines__P"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Safety_Considerations_Guidelines__P">Safety Considerations: Guidelines & Precautions (Appendix 2)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_nfkb_References"><a class="toc-item" href="/books/n/assayguide/nfkb/#nfkb.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons"><a class="toc-item" href="/books/n/assayguide/hcsneurons/">High Content Screening with Primary Neurons</a><div>Hassan Al-Ali, Murray Blackmore, John L Bixby, and Vance P. Lemmon.</div><div class="small">Published October 15, 2013; Last Update: October 1, 2014.</div><ul id="toc_lst_NBK53196_toc_hcsneurons" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Abstract"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Introduction"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Introduction">1. Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Prelminary_Assay_consideratio"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Prelminary_Assay_consideratio">2. Prelminary Assay considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Assay_Development"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Assay_Development">3. Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Additional_Considerations"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Additional_Considerations">4. Additional Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Protocols_for_HCS_with_Primar"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Protocols_for_HCS_with_Primar">5. Protocols for HCS with Primary Neurons</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_Acknowledgments"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.Acknowledgments">Acknowledgments</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_hcsneurons_References"><a class="toc-item" href="/books/n/assayguide/hcsneurons/#hcsneurons.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho"><a class="toc-item" href="/books/n/assayguide/phospho/">Phospho-ERK Assays</a><div>Authors: Kim E. Garbison, Beverly A. Heinz, Mary E. Lajiness, Jeffrey R. Weidner, and G. Sitta Sittampalam.</div><div class="small">Published May 1, 2012; Last Revision: June 25, 2015.</div><ul id="toc_lst_NBK53196_toc_phospho" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Abstract"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Introduction"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Overview_of_Technology"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Overview_of_Technology">Overview of Technology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_AlphaScreen_SureFire_ERK_Assay"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.AlphaScreen_SureFire_ERK_Assay">AlphaScreen SureFire ERK Assay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Characteristics_of_the_AlphaScre"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Characteristics_of_the_AlphaScre">Characteristics of the AlphaScreen SureFire ERK Assay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Sample_Protocol"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Sample_Protocol">Sample Protocol</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Assay_Formats"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Assay_Formats">Assay Formats</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Assay_Optimization"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Assay_Optimization">Assay Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Helpful_Hints_for_Performing_Sur"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Helpful_Hints_for_Performing_Sur">Helpful Hints for Performing SureFire AlphaScreen ERK Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_Websites"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.Websites">Websites</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_phospho_References"><a class="toc-item" href="/books/n/assayguide/phospho/#phospho.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3"><a class="toc-item" href="/books/n/assayguide/ip3/">IP-3/IP-1 Assays</a><div>Kim E. Garbison, Beverly A. Heinz, and Mary E. Lajiness.</div><div class="small">Published May 1, 2012.</div><ul id="toc_lst_NBK53196_toc_ip3" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Abstract"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Introduction"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Overview_of_Technology"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Overview_of_Technology">Overview of Technology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_IPOne_HTRF__Technology_Cisbio"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.IPOne_HTRF__Technology_Cisbio">IP-One HTRF<sup>®</sup> Technology (Cisbio)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Sample_Preparation_Protocol"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Sample_Preparation_Protocol">Sample Preparation Protocol</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Results_and_Data_Analysis"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Results_and_Data_Analysis">Results and Data Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Assay_Formats"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Assay_Formats">Assay Formats</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Assay_Optimization"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Assay_Optimization">Assay Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_Web_Sites"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.Web_Sites">Web Sites</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ip3_References"><a class="toc-item" href="/books/n/assayguide/ip3/#ip3.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/">Cardiomyocyte Impedance Assays</a><div>Sarah D. Lamore, Clay W Scott, and Matthew F. Peters.</div><div class="small">Published February 25, 2015.</div><ul id="toc_lst_NBK53196_toc_cardiomyocyte" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Abstract"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Flowchart"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Introduction"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Concept_and_Overview_of_xC"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Concept_and_Overview_of_xC">Concept and Overview of xCELLigence RTCA Cardio System</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Sample_Protocol"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Sample_Protocol">Sample Protocol</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_Assay_Optimization"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.Assay_Optimization">Assay Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cardiomyocyte_References"><a class="toc-item" href="/books/n/assayguide/cardiomyocyte/#cardiomyocyte.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa"><a class="toc-item" href="/books/n/assayguide/cetsa/">Screening for Target Engagement using the Cellular Thermal Shift Assay - CETSA</a><div>Hanna Axelsson, Helena Almqvist, Brinton Seashore-Ludlow, and Thomas Lundbäck.</div><div class="small">Published July 1, 2016.</div><ul id="toc_lst_NBK53196_toc_cetsa" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Abstract"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Flow_Chart"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Flow_Chart">Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Introduction"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Assay_Design"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Assay_Design">Assay Design</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Assay_Development"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Assay_Development">Assay Development</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Assay_Validation"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Assay_Validation">Assay Validation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Materials_and_Reagents"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Materials_and_Reagents">Materials and Reagents</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Reduction_of_Assay_Variability_and"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Reduction_of_Assay_Variability_and">Reduction of Assay Variability and Troubleshooting</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_Tentative_Applications"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.Tentative_Applications">Tentative Applications</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_cetsa_References"><a class="toc-item" href="/books/n/assayguide/cetsa/#cetsa.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp"><a class="toc-item" href="/books/n/assayguide/camp/">Measurement of cAMP for G<sub>αs</sub>- and G<sub>αi</sub> Protein-Coupled Receptors (GPCRs)</a><div>Tao Wang, Zhuyin Li, Mary Ellen Cvijic, Litao Zhang, and Chi Shing Sum.</div><div class="small">Published November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_camp" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Introduction"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Flowchart_of_Assay_Development_Guid"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Flowchart_of_Assay_Development_Guid">Flowchart of Assay Development Guidelines</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Overview_of_the_Assay_Technology"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Overview_of_the_Assay_Technology">Overview of the Assay Technology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Importance_of_cAMP_Standard_Curve"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Importance_of_cAMP_Standard_Curve">Importance of cAMP Standard Curve</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Assay_Development_and_Optimization"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Assay_Development_and_Optimization">Assay Development and Optimization Guidelines</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_cAMP_Assays_for_Receptors_Coupled_t"><a class="toc-item" href="/books/n/assayguide/camp/#camp.cAMP_Assays_for_Receptors_Coupled_t">cAMP Assays for Receptors Coupled to G<sub>αs</sub> Protein</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_cAMP_Assays_for_Receptors_Coupled_t_1"><a class="toc-item" href="/books/n/assayguide/camp/#camp.cAMP_Assays_for_Receptors_Coupled_t_1">cAMP Assays for Receptors Coupled to G<sub>αi</sub> Protein</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/camp/#camp.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_camp_References"><a class="toc-item" href="/books/n/assayguide/camp/#camp.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/">Measurement of β-Arrestin Recruitment for GPCR Targets</a><div>Tao Wang, Zhuyin Li, Mary Ellen Cvijic, Carol Krause, Litao Zhang, and Chi Shing Sum.</div><div class="small">Published November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_beta_arrestin" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Introduction"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Flowchart_of_Assay_Develop"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Flowchart_of_Assay_Develop">Flowchart of Assay Development Guidelines</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Overview_of_the__Arrestin"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Overview_of_the__Arrestin">Overview of the β-Arrestin Recruitment Assay</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Guidelines_and_Sample_Prot"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Guidelines_and_Sample_Prot">Guidelines and Sample Protocol for Agonist Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Guidelines_and_Sample_Prot_1"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Guidelines_and_Sample_Prot_1">Guidelines and Sample Protocol for Antagonist Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Troubleshooting"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Troubleshooting">Troubleshooting</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_beta_arrestin_References"><a class="toc-item" href="/books/n/assayguide/beta_arrestin/#beta_arrestin.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit"><a class="toc-item" href="/books/n/assayguide/gen_edit/">Genome Editing Using Engineered Nucleases and Their Use in Genomic Screening</a><div>Joana R. Costa, Bruce E. Bejcek, James E. McGee, Adam I. Fogel, Kyle R. Brimacombe, and Robin Ketteler.</div><div class="small">Published November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_gen_edit" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Abstract"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Introduction"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Screening"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Screening">Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Pooled_Library_Screening"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Pooled_Library_Screening">Pooled Library Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Arrayed_Library_Screening"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Arrayed_Library_Screening">Arrayed Library Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Conclusion"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Appendix"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Appendix">Appendix</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_References"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Suggested_Reading"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Suggested_Reading">Suggested Reading</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_gen_edit_Suggested_Websites"><a class="toc-item" href="/books/n/assayguide/gen_edit/#gen_edit.Suggested_Websites">Suggested Websites</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell"><a class="toc-item" href="/books/n/assayguide/ppi_cell/">Inhibition of Protein-Protein Interactions: Cell-Based Assays</a><div>Mark Wade, Jacqui Méndez, Nathan P. Coussens, Michelle R. Arkin, and Marcie A. Glicksman.</div><div class="small">Published November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_ppi_cell" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Abstract"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Introduction"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Background"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Background">Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_General_Considerations_for_Deve"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.General_Considerations_for_Deve">General Considerations for Developing Cell-Based PPI Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_F_rster_Resonance_Energy_Transf"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.F_rster_Resonance_Energy_Transf">Förster Resonance Energy Transfer (FRET)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Bioluminescence_Resonance_Energ"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Bioluminescence_Resonance_Energ">Bioluminescence Resonance Energy Transfer (BRET)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Fluorescent_Protein_Fragment_Co"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Fluorescent_Protein_Fragment_Co">Fluorescent Protein Fragment Complementation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Luciferase_Enzyme_Fragment_Comp"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Luciferase_Enzyme_Fragment_Comp">Luciferase Enzyme Fragment Complementation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Alternative_Enzyme_Complementat"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Alternative_Enzyme_Complementat">Alternative Enzyme Complementation Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_Suggested_Websites"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.Suggested_Websites">Suggested Websites</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ppi_cell_References"><a class="toc-item" href="/books/n/assayguide/ppi_cell/#ppi_cell.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr"><a class="toc-item" href="/books/n/assayguide/pcgpcr/">Pharmacological Characterization of GPCR Agonists, Antagonists, Allosteric Modulators and Biased Ligands from HTS Hits to Lead Optimization</a><div>Chi Shing Sum, Brian J. Murphy, Zhuyin Li, Tao Wang, Litao Zhang, and Mary Ellen Cvijic.</div><div class="small">Published November 1, 2019.</div><ul id="toc_lst_NBK53196_toc_pcgpcr" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Abstract"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Introduction_to_GPCRs_as_Drug_Tar"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Introduction_to_GPCRs_as_Drug_Tar">Introduction to GPCRs as Drug Targets</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_GPCR_Signaling_from_a_Drug_Discov"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.GPCR_Signaling_from_a_Drug_Discov">GPCR Signaling from a Drug Discovery Perspective</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_HighThroughput_Screening_Assays_f"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.HighThroughput_Screening_Assays_f">High-Throughput Screening Assays for Identification and Optimization of GPCR Ligands</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Pharmacological_Deliverables_of_H"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Pharmacological_Deliverables_of_H">Pharmacological Deliverables of HTS and Lead Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Important_Pharmacological_Conside"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Important_Pharmacological_Conside">Important Pharmacological Considerations During Lead Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_How_the_On_and_OffRates_of_the_Te"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.How_the_On_and_OffRates_of_the_Te">How the On- and Off-Rates of the Test Agonist (Compound Kinetics) Affect Compound Functional Activity Measurements and Thereby Impact Lead Optimization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_DoseShift_Assays"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.DoseShift_Assays">Dose-Shift Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_DoseShift_Assays_in_Screening_Mod"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.DoseShift_Assays_in_Screening_Mod">Dose-Shift Assays in Screening Mode</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Biased_Signaling"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Biased_Signaling">Biased Signaling</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Conclusion"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Conclusion">Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Acknowledgement"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Acknowledgement">Acknowledgement</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_Keywords"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.Keywords">Keywords</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pcgpcr_References"><a class="toc-item" href="/books/n/assayguide/pcgpcr/#pcgpcr.References">References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoassay_xml"><a class="toc-item" href="/books/n/assayguide/invivoassay.xml/">
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<i>In Vivo</i> Assay Guidelines</a><div>Editors: Viswanath Devanarayan and Philip W. Iversen.</div><ul id="toc_lst_NBK53196_toc_invivoassay_xml" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide"><a class="toc-item" href="/books/n/assayguide/invivoguide/"><i>In Vivo</i> Assay Guidelines</a><div>Joseph Haas, Jason Manro, Harlan Shannon, Wes Anderson, Joe Brozinick, Arunava Chakravartty, Mark Chambers, Jian Du, Brian Eastwood, Joe Heuer, Stephen Iturria, Philip Iversen, Dwayne Johnson, Kirk Johnson, Michael O’Neill, Hui-Rong Qian, Dana Sindelar, and Kjell Svensson.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_invivoguide" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_Flow_Chart"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.Flow_Chart">Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_1_Introduction"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.1_Introduction">1. Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_2_Assay_Validation_Procedure"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.2_Assay_Validation_Procedure">2. Assay Validation Procedures</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_3_Design_Sample_Size_Randomi"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.3_Design_Sample_Size_Randomi">3. Design, Sample Size, Randomization, and Analysis Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_4_Abbreviations"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.4_Abbreviations">4. Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_invivoguide_5_Suggested_Reading"><a class="toc-item" href="/books/n/assayguide/invivoguide/#invivoguide.5_Suggested_Reading">5. Suggested Reading</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror"><a class="toc-item" href="/books/n/assayguide/ivror/"><i>In Vivo</i> Receptor Occupancy in Rodents by LC-MS/MS</a><div>Cynthia D. Jesudason, Susan DuBois, Megan Johnson, Vanessa N. Barth, and Anne B. Need.</div><div class="small">Published March 9, 2017.</div><ul id="toc_lst_NBK53196_toc_ivror" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror_Abstract"><a class="toc-item" href="/books/n/assayguide/ivror/#ivror.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror_Experimental_Design"><a class="toc-item" href="/books/n/assayguide/ivror/#ivror.Experimental_Design">Experimental Design</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror_Flowchart"><a class="toc-item" href="/books/n/assayguide/ivror/#ivror.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror_Materials_and_Methods"><a class="toc-item" href="/books/n/assayguide/ivror/#ivror.Materials_and_Methods">Materials and Methods</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ivror_References"><a class="toc-item" href="/books/n/assayguide/ivror/#ivror.References">References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_artifacts"><a class="toc-item" href="/books/n/assayguide/artifacts/">Assay Artifacts and Interferences</a><div>Editors: Nathan P. Coussens and Jayme L. Dahlin.</div><ul id="toc_lst_NBK53196_toc_artifacts" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference"><a class="toc-item" href="/books/n/assayguide/assayinterference/">Compound-Mediated Assay Interferences in Homogeneous Proximity Assays</a><div>Nathan P. Coussens, Douglas Auld, Philippe Roby, Jarrod Walsh, Jonathan B. Baell, Stephen Kales, Kamyar Hadian, and Jayme L. Dahlin.</div><div class="small">Published February 1, 2020.</div><ul id="toc_lst_NBK53196_toc_assayinterference" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Abstract"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Flowchart"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Abbreviations"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Introduction_to_Proxim"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Introduction_to_Proxim">Introduction to Proximity Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Chemical_Mechanisms_of"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Chemical_Mechanisms_of">Chemical Mechanisms of Assay Interference in Homogeneous Proximity Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Strategies_to_Identify"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Strategies_to_Identify">Strategies to Identify Compound-Mediated Interference in Homogeneous Proximity Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Strategies_to_Mitigate"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Strategies_to_Mitigate">Strategies to Mitigate Compound-Mediated Interferences in Proximity Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Conclusions"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Suggested_Readings_alp"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Suggested_Readings_alp">Suggested Readings (alphabetical order)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Suggested_Web_Resource"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Suggested_Web_Resource">Suggested Web Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Glossary"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Glossary">Glossary</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_Conflict_of_Interest_S"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.Conflict_of_Interest_S">Conflict of Interest Statement</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayinterference_References"><a class="toc-item" href="/books/n/assayguide/assayinterference/#assayinterference.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact"><a class="toc-item" href="/books/n/assayguide/chemreact/">Assay Interference by Chemical Reactivity</a><div>Jayme L. Dahlin, Jonathan Baell, and Michael A. Walters.</div><div class="small">Published September 18, 2015.</div><ul id="toc_lst_NBK53196_toc_chemreact" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Abstract"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Flow_Chart"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Flow_Chart">Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Abbreviations"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Introduction_and_Background"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Introduction_and_Background">Introduction and Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_KnowledgeBased_Strategies_to_M"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.KnowledgeBased_Strategies_to_M">Knowledge-Based Strategies to Minimize Impact of Interference Compounds</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_ExperimentalBased_Strategies_t"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.ExperimentalBased_Strategies_t">Experimental-Based Strategies to Mitigate the Impact of Reactive Interference Compounds</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Conclusions"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Suggested_Websites"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Suggested_Websites">Suggested Websites</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_Suggested_Readings_alphabetica"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.Suggested_Readings_alphabetica">Suggested Readings (alphabetical order)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_chemreact_References"><a class="toc-item" href="/books/n/assayguide/chemreact/#chemreact.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa"><a class="toc-item" href="/books/n/assayguide/interferefa/">Interference with Fluorescence and Absorbance</a><div>Anton Simeonov and Mindy I. Davis.</div><div class="small">Published December 7, 2015; Last Update: July 1, 2018.</div><ul id="toc_lst_NBK53196_toc_interferefa" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_Abstract"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_Flow_Chart"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.Flow_Chart">Flow Chart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_Fluorescence_Interferences"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.Fluorescence_Interferences">Fluorescence Interferences</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_Absorbance_Interferences"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.Absorbance_Interferences">Absorbance Interferences</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_interferefa_References"><a class="toc-item" href="/books/n/assayguide/interferefa/#interferefa.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre"><a class="toc-item" href="/books/n/assayguide/ilre/">Interferences with Luciferase Reporter Enzymes</a><div>Douglas S. Auld and James Inglese.</div><div class="small">Published July 1, 2016; Last Update: July 1, 2018.</div><ul id="toc_lst_NBK53196_toc_ilre" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre_Abstract"><a class="toc-item" href="/books/n/assayguide/ilre/#ilre.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre_Introduction"><a class="toc-item" href="/books/n/assayguide/ilre/#ilre.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre_Interferences_with_Reporter_Enzymes"><a class="toc-item" href="/books/n/assayguide/ilre/#ilre.Interferences_with_Reporter_Enzymes">Interferences with Reporter Enzymes</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre_Strategies_to_Mitigate_Luciferase_I"><a class="toc-item" href="/books/n/assayguide/ilre/#ilre.Strategies_to_Mitigate_Luciferase_I">Strategies to Mitigate Luciferase Inhibitor Interference</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ilre_References"><a class="toc-item" href="/books/n/assayguide/ilre/#ilre.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation"><a class="toc-item" href="/books/n/assayguide/aggregation/">Assay Interference by Aggregation</a><div>Douglas S. Auld, James Inglese, and Jayme L. Dahlin.</div><div class="small">Published July 26, 2017.</div><ul id="toc_lst_NBK53196_toc_aggregation" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Abstract"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Flowchart"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Flowchart">Flowchart</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Abbreviations"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Introduction_and_Background"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Introduction_and_Background">Introduction and Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Strategies_to_Mitigate_Incid"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Strategies_to_Mitigate_Incid">Strategies to Mitigate Incidence of Aggregation in Biochemical Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Strategies_to_Identify_Aggre"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Strategies_to_Identify_Aggre">Strategies to Identify Aggregation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Frequently_Asked_Questions"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Frequently_Asked_Questions">Frequently Asked Questions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Conclusions"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Suggested_Web_Resources"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Suggested_Web_Resources">Suggested Web Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Suggested_Readings_alphabeti"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Suggested_Readings_alphabeti">Suggested Readings (alphabetical order)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_aggregation_REFERENCES"><a class="toc-item" href="/books/n/assayguide/aggregation/#aggregation.REFERENCES">REFERENCES</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_avoqc"><a class="toc-item" href="/books/n/assayguide/avoqc/">Assay Validation, Operations and Quality Control</a><div>Editors: Viswanath Devanarayan, Philip W. Iversen, Jeffrey R. Weidner, and Mary Jo Wildey.</div><ul id="toc_lst_NBK53196_toc_avoqc" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting"><a class="toc-item" href="/books/n/assayguide/datareporting/">Basic Guidelines for Reporting Non-Clinical Data</a><div>Jayme L. Dahlin, G. Sitta Sittampalam, Nathan P. Coussens, Viswanath Devanarayan, Jeffrey R. Weidner, Philip W. Iversen, Joseph V. Haas, Duane D. Bronson, O. Joseph Trask, Jr., Todd R. Wiernicki, and Steven D. Kahl.</div><div class="small">Published October 1, 2019.</div><ul id="toc_lst_NBK53196_toc_datareporting" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Abstract"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Abbreviations"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Abbreviations">Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Overview"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Overview">Overview</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Rounding_to_Decimal_Places"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Rounding_to_Decimal_Places">Rounding to Decimal Places</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Rounding_to_Significant_Di"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Rounding_to_Significant_Di">Rounding to Significant Digits</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Presenting_Rounded_Data_in"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Presenting_Rounded_Data_in">Presenting Rounded Data in Tables</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Logarithms"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Logarithms">Logarithms</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Statistical_DescriptorsMet"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Statistical_DescriptorsMet">Statistical Descriptors/Metrics</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Example_of_Methods_to_Plot"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Example_of_Methods_to_Plot">Example of Methods to Plot the Same Data</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Correlation_of_Data"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Correlation_of_Data">Correlation of Data</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Agreement_Between_Two_Vari"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Agreement_Between_Two_Vari">Agreement Between Two Variables</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Pvalues"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Pvalues">P-values</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Calculation_of_Upper_and_L"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Calculation_of_Upper_and_L">Calculation of Upper and Lower Quantitation Limits</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Area_under_the_Curve"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Area_under_the_Curve">Area under the Curve</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Graph_Elements"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Graph_Elements">Graph Elements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Types_of_Graphs"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Types_of_Graphs">Types of Graphs</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_GraphingStatistical_Softwa"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.GraphingStatistical_Softwa">Graphing/Statistical Software Programs</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Tables"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Tables">Tables</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Interactive_Data_Visualiza"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Interactive_Data_Visualiza">Interactive Data Visualization</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_Databases"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.Databases">Databases</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_datareporting_References"><a class="toc-item" href="/books/n/assayguide/datareporting/#datareporting.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap"><a class="toc-item" href="/books/n/assayguide/baodevap/">Development and Applications of the Bioassay Ontology (BAO) to Describe and Categorize High-Throughput Assays</a><div>Uma D. Vempati and Stephan C. Schürer.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_baodevap" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Abstract"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Introduction"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_The_BAO_to_organize_screening_a"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.The_BAO_to_organize_screening_a">The BAO to organize screening assays, results and campaigns</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Ontology_development_and_overvi"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Ontology_development_and_overvi">Ontology development and overview of the BAO</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Application_of_the_BAO"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Application_of_the_BAO">Application of the BAO</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Assay_Annotation_Tool_to_facili"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Assay_Annotation_Tool_to_facili">Assay Annotation Tool to facilitate adaptation of the BAO</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_BAOSearch_software_application"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.BAOSearch_software_application">BAOSearch software application</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Summary_and_conclusion"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Summary_and_conclusion">Summary and conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_References"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.References">References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_baodevap_Glossary_of_BAO_terms_partial_l"><a class="toc-item" href="/books/n/assayguide/baodevap/#baodevap.Glossary_of_BAO_terms_partial_l">Glossary of BAO terms (partial list)</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm"><a class="toc-item" href="/books/n/assayguide/dsrm/">Data Standardization for Results Management</a><div>Robert M. Campbell, Julia Dymshitz, Brian J. Eastwood, Renee Emkey, David P. Greenen, Julia M. Heerding, Dwayne Johnson, Thomas H. Large, Thomas Littlejohn, Chahrzad Montrose, Suzanne E. Nutter, Barry D. Sawyer, Sandra K. Sigmund, Martin Smith, Jeffrey R. Weidner, and Richard W. Zink.</div><div class="small">Published May 1, 2012; Last Update: October 20, 2017.</div><ul id="toc_lst_NBK53196_toc_dsrm" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Abstract"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Introduction"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Data_Types_and_Associated_Rules_for"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Data_Types_and_Associated_Rules_for">Data Types and Associated Rules for Radioligand Binding Assays: Inhibition Mode</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Data_Types_and_Associated_Rules_for_1"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Data_Types_and_Associated_Rules_for_1">Data Types and Associated Rules for Enzymatic Assays: Inhibition Mode</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Data_Types_and_Associated_Rules_for_2"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Data_Types_and_Associated_Rules_for_2">Data Types and Associated Rules for <i>In Vitro</i> Functional Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Reference"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Reference">Reference</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_dsrm_Glossary"><a class="toc-item" href="/books/n/assayguide/dsrm/#dsrm.Glossary">Glossary</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation"><a class="toc-item" href="/books/n/assayguide/htsvalidation/">HTS Assay Validation</a><div>Philip W. Iversen, Benoit Beck, Yun-Fei Chen, Walthere Dere, Viswanath Devanarayan, Brian J Eastwood, Mark W. Farmen, Stephen J. Iturria, Chahrzad Montrose, Roger A. Moore, Jeffrey R. Weidner, and G. Sitta Sittampalam.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_htsvalidation" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_Abstract"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_Overview"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.Overview">1. Overview</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_Stability_and_Process_Stud"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.Stability_and_Process_Stud">2. Stability and Process Studies</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_Plate_Uniformity_and_Signa"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.Plate_Uniformity_and_Signa">3. Plate Uniformity and Signal Variability Assessment</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_ReplicateExperiment_Study"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.ReplicateExperiment_Study">4. Replicate-Experiment Study</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_How_to_Deal_with_High_Assa"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.How_to_Deal_with_High_Assa">5. How to Deal with High Assay Variability</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_Bridging_Studies_for_Assay"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.Bridging_Studies_for_Assay">6. Bridging Studies for Assay Upgrades and Minor Changes</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_htsvalidation_References"><a class="toc-item" href="/books/n/assayguide/htsvalidation/#htsvalidation.References">7. References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayops"><a class="toc-item" href="/books/n/assayguide/assayops/">Assay Operations for SAR Support</a><div>Benoit Beck, Yun-Fei Chen, Walthere Dere, Viswanath Devanarayan, Brian J. Eastwood, Mark W. Farmen, Stephen J. Iturria, Phillip W. Iversen, Steven D. Kahl, Roger A. Moore, Barry D. Sawyer, and Jeffrey Weidner.</div><div class="small">Published May 1, 2012; Last Update: November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_assayops" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayops_Abstract"><a class="toc-item" href="/books/n/assayguide/assayops/#assayops.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayops_Determination_of_EC50IC50"><a class="toc-item" href="/books/n/assayguide/assayops/#assayops.Determination_of_EC50IC50">A. Determination of EC<sub>50</sub>/IC<sub>50</sub></a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayops_Production_Monitoring"><a class="toc-item" href="/books/n/assayguide/assayops/#assayops.Production_Monitoring">B. Production Monitoring</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayops_References"><a class="toc-item" href="/books/n/assayguide/assayops/#assayops.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio"><a class="toc-item" href="/books/n/assayguide/minsigratio/">Minimum Significant Ratio – A Statistic to Assess Assay Variability</a><div>Joseph V. Haas, Brian J. Eastwood, Philip W. Iversen, Viswanath Devanarayan, and Jeffrey R. Weidner.</div><div class="small">Published November 1, 2013; Last Update: November 20, 2017.</div><ul id="toc_lst_NBK53196_toc_minsigratio" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_Abstract"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_sec1"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.sec1">1. Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_sec2"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.sec2">2. Common Types of MSR</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_sec3"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.sec3">3. Other Considerations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_sec4"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.sec4">4. Conclusion</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_minsigratio_sec5"><a class="toc-item" href="/books/n/assayguide/minsigratio/#minsigratio.sec5">5. References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assaytechs"><a class="toc-item" href="/books/n/assayguide/assaytechs/">Assay Technologies</a><div>Editors: Marcie Glicksman, James McGee, Owen McManus, and G. Sitta Sittampalam.</div><ul id="toc_lst_NBK53196_toc_assaytechs" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec"><a class="toc-item" href="/books/n/assayguide/massspec/">HPLC-MS/MS for Hit Generation</a><div>Stefan Jon Thibodeaux, David A Yurek, and James E McGee.</div><div class="small">Last Update: July 8, 2017.</div><ul id="toc_lst_NBK53196_toc_massspec" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Abstract"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Introduction"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Chromatography"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Chromatography">Chromatography</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Instrument_Set_Up"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Instrument_Set_Up">Instrument Set Up</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Data_Handling"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Data_Handling">Data Handling</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_Other_Notes"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.Other_Notes">Other Notes</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_massspec_References"><a class="toc-item" href="/books/n/assayguide/massspec/#massspec.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt"><a class="toc-item" href="/books/n/assayguide/ibt/">Impedance-Based Technologies</a><div>Kim E. Garbison, Beverly A. Heinz, Mary E. Lajiness, Jeffrey R. Weidner, and G. Sitta Sittampalam.</div><div class="small">Published May 1, 2012.</div><ul id="toc_lst_NBK53196_toc_ibt" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Abstract"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Introduction"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Overview_of_ACEA_Biosciences_RTCES"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Overview_of_ACEA_Biosciences_RTCES">Overview of ACEA Biosciences RT-CES<sup>®</sup></a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Overview_of_CellKeyTM_System"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Overview_of_CellKeyTM_System">Overview of CellKey<sup>TM</sup> System</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Sample_Protocol_for_CellKeyTM_MDS_An"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Sample_Protocol_for_CellKeyTM_MDS_An">Sample Protocol for CellKey<sup>TM</sup> (MDS Analytical Technologies)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_CellKeyTM_System_Results_and_Data_An"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.CellKeyTM_System_Results_and_Data_An">CellKey<sup>TM</sup> System Results and Data Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Helpful_Hints_for_Performing_CellKey"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Helpful_Hints_for_Performing_CellKey">Helpful Hints for Performing CellKey<sup>TM</sup> Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Additional_References"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Additional_References">Additional References</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_ibt_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/ibt/#ibt.Acknowledgements">Acknowledgements</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_instrumentation"><a class="toc-item" href="/books/n/assayguide/instrumentation/">Instrumentation</a><div>Editors: Nathan P. Coussens, Sarine Markossian, and G. Sitta Sittampalam.</div><ul id="toc_lst_NBK53196_toc_instrumentation" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates"><a class="toc-item" href="/books/n/assayguide/microplates/">Microplate Selection and Recommended Practices in High-throughput Screening and Quantitative Biology</a><div>Douglas S. Auld, Ph.D., Peter A. Coassin, B.S., Nathan P. Coussens, Ph.D., Paul Hensley, Carleen Klumpp-Thomas, Sam Michael, G. Sitta Sittampalam, Ph.D., O. Joseph Trask, B.S., Bridget K. Wagner, Ph.D., Jeffrey R. Weidner, Ph.D., Mary Jo Wildey, Ph.D., and Jayme L. Dahlin, M.D., Ph.D.</div><div class="small">Published June 1, 2020.</div><ul id="toc_lst_NBK53196_toc_microplates" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Abstract"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Key_Abbreviations"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Key_Abbreviations">Key Abbreviations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Introduction_to_Microplates"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Introduction_to_Microplates">Introduction to Microplates in High-Throughput Screening and Quantitative Biology</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Technical_Factors_in_Micropl"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Technical_Factors_in_Micropl">Technical Factors in Microplate Selection</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_TechnologySpecific_Considera"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.TechnologySpecific_Considera">Technology-Specific Considerations – High-Content Screening</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Microplate_Usage"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Microplate_Usage">Microplate Usage</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Troubleshooting_and_Special"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Troubleshooting_and_Special">Troubleshooting and Special Topics</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Conclusions"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Conclusions">Conclusions</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Suggested_Readings_alphabeti"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Suggested_Readings_alphabeti">Suggested Readings (alphabetical order)</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Glossary"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Glossary">Glossary</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Acknowledgements"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Acknowledgements">Acknowledgements</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_Conflict_of_Interest_Stateme"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.Conflict_of_Interest_Stateme">Conflict of Interest Statement</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_microplates_References"><a class="toc-item" href="/books/n/assayguide/microplates/#microplates.References">References</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip"><a class="toc-item" href="/books/n/assayguide/assayequip/">Basics of Assay Equipment and Instrumentation for High Throughput Screening</a><div>Eric Jones, Sam Michael, and G. Sitta Sittampalam.</div><div class="small">Published May 1, 2012; Last Update: April 2, 2016.</div><ul id="toc_lst_NBK53196_toc_assayequip" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Abstract"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Introduction"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Common_Equipment_in_HTS_Labs"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Common_Equipment_in_HTS_Labs">Common Equipment in HTS Labs</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Microplates"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Microplates">Microplates</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Microplate_Sealing"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Microplate_Sealing">Microplate Sealing</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Microplate_Readers"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Microplate_Readers">Microplate Readers</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Spectrophotometry"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Spectrophotometry">Spectrophotometry</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_pH_Meters"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.pH_Meters">pH Meters</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Electronic_Balances"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Electronic_Balances">Electronic Balances</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Microscopes"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Microscopes">Microscopes</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Liquid_Handling_Devices"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Liquid_Handling_Devices">Liquid Handling Devices</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Suggested_Websites_and_Resour"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Suggested_Websites_and_Resour">Suggested Websites and Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_assayequip_Suggested_Readings_alphabetic"><a class="toc-item" href="/books/n/assayguide/assayequip/#assayequip.Suggested_Readings_alphabetic">Suggested Readings (alphabetical order)</a></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc"><a class="toc-item" href="/books/n/assayguide/radiocalc/">Calculations and Instrumentation used for Radioligand Binding Assays</a><div>Steven D. Kahl, G. Sitta Sittampalam, and Jeffrey Weidner.</div><div class="small">Published May 1, 2012; Last Update: October 1, 2012.</div><ul id="toc_lst_NBK53196_toc_radiocalc" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Abstract"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Introduction"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Introduction">Introduction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Radioactive_Calculations"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Radioactive_Calculations">Radioactive Calculations</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Instrumentation"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Instrumentation">Instrumentation</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Uniformity_Plate"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Uniformity_Plate">Uniformity Plate</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Color_Quench_Correction"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Color_Quench_Correction">Color Quench Correction</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_radiocalc_Abbreviations"><a class="toc-item" href="/books/n/assayguide/radiocalc/#radiocalc.Abbreviations">Abbreviations</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_pharmadm"><a class="toc-item" href="/books/n/assayguide/pharmadm/">Pharmacokinetics and Drug Metabolism</a><div>Editors: Thomas D.Y. Chung and Xin Xu.</div><ul id="toc_lst_NBK53196_toc_pharmadm" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk"><a class="toc-item" href="/books/n/assayguide/admepk/"><i>In Vitro</i> and <i>In Vivo</i> Assessment of ADME and PK Properties During Lead Selection and Lead Optimization – Guidelines, Benchmarks and Rules of Thumb</a><div>Thomas D.Y. Chung, David B. Terry, and Layton H. Smith.</div><div class="small">Published September 9, 2015.</div><ul id="toc_lst_NBK53196_toc_admepk" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_Abstract"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_Flow_Chart_of_a_Twotier_Approach"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.Flow_Chart_of_a_Twotier_Approach">Flow Chart of a Two-tier Approach for <i>In Vitro</i> and <i>In Vivo</i> Analysis</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_Background"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.Background">Background</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_In_Vitro_Analysis__Low_Compound_R"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.In_Vitro_Analysis__Low_Compound_R"><i>In Vitro</i> Analysis - Low Compound Requirements and Relative Moderate Capacity</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_Example_of_In_Vitro_ADME_Profilin"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.Example_of_In_Vitro_ADME_Profilin">Example of <i>In Vitro</i> ADME Profiling Assays</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_In_Vivo_Analysis__High_Compound_R"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.In_Vivo_Analysis__High_Compound_R"><i>In Vivo</i> Analysis - High Compound Requirements and Low Capacity</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_Suggested_Equipment_and_Resources"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.Suggested_Equipment_and_Resources">Suggested Equipment and Resources</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_admepk_References"><a class="toc-item" href="/books/n/assayguide/admepk/#admepk.References">References</a></li></ul></li></ul></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_glossary_xml"><a class="toc-item" href="/books/n/assayguide/glossary.xml/">Glossary</a><div>Editors: Nathan P. Coussens, Sarine Markossian, G. Sitta Sittampalam, and Jeffrey R. Weidner.</div><ul id="toc_lst_NBK53196_toc_glossary_xml" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_qbiogloss"><a class="toc-item" href="/books/n/assayguide/qbiogloss/">Glossary of Quantitative Biology Terms</a><div>Viswanath Devanarayan, Barry D. Sawyer, Chahrzad Montrose, Dwayne Johnson, David P. Greenen, Sitta Sittampalam, Terry Riss, and Lisa Minor.</div><div class="small">Published April 15, 2012; Last Update: September 22, 2014.</div><ul id="toc_lst_NBK53196_toc_qbiogloss" class="simple-list toc bktoc_lst_exp"><li class="half_rhythm" id="toc_itm_NBK53196_toc_qbiogloss_Abstract"><a class="toc-item" href="/books/n/assayguide/qbiogloss/#qbiogloss.Abstract">Abstract</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_qbiogloss_Cell_Culture_Terms"><a class="toc-item" href="/books/n/assayguide/qbiogloss/#qbiogloss.Cell_Culture_Terms">Cell Culture Terms</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_qbiogloss_Quantitative_Biology_Terms"><a class="toc-item" href="/books/n/assayguide/qbiogloss/#qbiogloss.Quantitative_Biology_Terms">Quantitative Biology Terms</a></li><li class="half_rhythm" id="toc_itm_NBK53196_toc_qbiogloss_Genetics_Terms"><a class="toc-item" href="/books/n/assayguide/qbiogloss/#qbiogloss.Genetics_Terms">Genetics Terms</a></li></ul></li></ul></li></ul><div class="bktoc_all_cntnr align_right" style="display:none"><ul class="inline_list_right"><li><a class="bktoc_all_exp" href="#">Expand All</a></li><li style="margin-left:.8em"><a class="bktoc_all_clps" href="#">Collapse All</a></li></ul></div></div><div><p>Previous Editors: Bruce Bejcek, Jose M.M. Caaveiro, Neely Gal-Edd, Steven D. Kahl, Stephen C. Kales, Susan Kirshner, Vance P. Lemmon, Lisa Minor, Andrew Napper, Henrike Nelsen, John M. Peltier, Yueming Wang</p><p>Contact Information: <a href="mailto:dev@null" data-email="vog.hin@naissokram.eniras" class="oemail">vog.hin@naissokram.eniras</a>
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