High Throughput Screening Methods and Protocols

This third edition volume expands on the previous editions both by presenting more detailed protocols for the techniques described in the first and second editions of High Throughput Screening: Methods and Protocols and by covering important new procedures. The first chapter of this book provides an overview of important assay development techniques, while the rest of the chapters detail how to develop and execute screens at whatever throughput the user needs. Some chapter examples are: structure-based virtual screening,  high throughput screening using mass spectrometry, identification of state-dependent blockers for voltage gated calcium channels, bioluminescence resonance energy transfer platform to monitor protein-protein interactions in live cells, high throughput flow cytometry, and application of imaging-based assays in microplate formats for high content screening. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting edge and thorough, High Throughput Screening: Methods and Protocols, Third Edition, is a valuable resource for anyone who is interested in HTS research.

1. Design and Implementation of High Throughput Screening Assays
David J. Powell, Robert P. Hertsberg, and Ricardo Macarr?n

2. Characterization of Inhibitor Binding Through Multiple Inhibitor Analysis: A Novel Local Fitting Method
Thomas V. Riera, Tim J. Wigle, and Robert A. Copeland

3. High Throughput Screening Using Mass Spectrometry within Drug Discovery
Mattias Rohman and Jonathan Wingfield

4. Structure-Based Virtual Screening of Commercially Available Compound Libraries
Dmitri Kireev

5. AlphaScreen-Based Assays: Ultra-High Throughput Screening for Small Molecule Inhibitors of Challenging Enzymes and Protein-Protein Interactions
Adam Yasgar, Ajit Jadhav, Anton Simeonov, and Nathan P. Coussens

6. Instrument Quality Control
Chatura Jayakody and Emily A. Hull-Ryde

7. Application of Fluorescence Polarization in HTS Assays< Xinyi Huang and Ann Aulabaugh

8. Time Resolved Fluorescence Assays
Chen-Ting Ma and Eduard A. Sergienko

9. Protein Kinase Selectivity Profiling Using Microfluid Mobility Shift Assays
Peter Drueckes

10. Screening for Inhibitors of Kinase Autophosphorylation
Bianca Heedmann and Martin Klumpp

11. A Fluorescence-Based High Throughput Screening Assay to Identify Growth Inhibitors of the Pathogenic Fungus Aspergillus fumigatus
Thomas M. Smith, Daryl L. Richie, Jianshi Tao

12. Mycobacterium tuberculosis High Throughput Screening
E. Lucile White, Nichole A. Tower, and Lynn Rasmussen

13. Identification of State-Dependent Blockers for Voltage Gated Calcium Channels Using a FLIPR Based Assay
Alberto di Silvio

14. A Luciferase Reporter Gene System for High Throughput Screening of ?-Globin Gene Activators
Wensheng Xie, Robert Silvers, Michael Ouellette, Zining Wu, Quinn Lu, Hu Li, Kathleen Gallagher, Kathy Johnson, and Monica Montoute

15. A High Throughput Flow Cytometry Assay for Identification of Inhibitors of 3’, 5’-Cyclic Adenosine Monophosphate Efflux
Dominique Perez, Peter C. Simons, Yelena Smagley, Larry A. Sklar, and Alexandre Chigaev

16. High Throughput Cell Toxicity Assays
David Murray, Lisa McWilliams, and Mark Wigglesworth

17. BRETⁿ: NanoLuc-Based Bioluminescence Resonance Energy Transfer Platform to Monitor Protein-Protein Interactions in Live Cells
Xiu-Lei Mo and Haian Fu

18. Application of Imaging-Based Assays in Microplate Formats for High Content Screening
Adam I. Fogel, Scott E. Martin, and Samuel A. Hasson