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, chaptersinclude 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.

Design and Implementation of High Throughput Screening Assays.- Characterization of Inhibitor Binding Through Multiple Inhibitor Analysis: A Novel Local Fitting Method.- High Throughput Screening Using Mass Spectrometry within Drug Discovery.- Structure-Based Virtual Screening of Commercially Available Compound Libraries.- AlphaScreen-Based Assays: Ultra-High Throughput Screening for Small Molecule Inhibitors of Challenging Enzymes and Protein-Protein Interactions.- Instrument Quality Control.- Application of Fluorescence Polarization in HTS Assays.- Time Resolved Fluorescence Assays.- Protein Kinase Selectivity Profiling Using Microfluid Mobility Shift Assays.- Screening for Inhibitors of Kinase Autophosphorylation.- A Fluorescence-Based High Throughput Screening Assay to Identify Growth Inhibitors of the Pathogenic Fungus Aspergillus fumigatus.- Mycobacterium tuberculosis High Throughput Screening.- Identification of State-Dependent Blockers for Voltage Gated Calcium Channels Using a FLIPR Based Assay.- A Luciferase Reporter Gene System for High Throughput Screening of ?-Globin Gene Activators.- A High Throughput Flow Cytometry Assay for Identification of Inhibitors of 3’, 5’-Cyclic Adenosine Monophosphate Efflux.- High Throughput Cell Toxicity Assays.- BRETⁿ: NanoLuc-Based Bioluminescence Resonance Energy Transfer Platform to Monitor Protein-Protein Interactions in Live Cells.- Application of Imaging-Based Assays in Microplate Formats for High Content Screening.