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Table of Contents
From Assay Guidance Wiki
Authors of Version 6
Assay Validation
- Overview
- Stability and Process Studies
- Plate Uniformity and Signal Variability Assessment
- Replicate-Experiment Study
- How to Deal With High Assay Variability
- Bridging Studies for Assay Upgrades and Minor Changes
- References
Assay Operations for SAR Support
Enzyme Assays
- Basic Enzyme Assays
- Overview of MOA in Drug Discovery
- Glossary of MOA Terms
- Types of Inhibition
- Performing MOA Studies
- Enzyme MOA References
Receptor Binding Assays
- Steps to Assay Development for SPA Format
- Steps to Assay Development for Filter Format
- Introduction
- Reagents
- Scintillation Proximity Assays (SPA)
- SPA Assay Format
- Assay Buffer
- Assay Conditions
- Binding Parameters
- Filtration Assays
- Filter Assay Format
- Assay Buffer
- Assay Conditions
- Binding Parameters
- Special Circumstances
- Practical Use of Fluorescence Polarization in Competitive Receptor Binding Assays
- Abbreviations
- Additional Resources
GTPγS Binding Assays
- Steps to Assay Development
- Introduction
- Materials and Reagents
- Membrane Preperations and Assay Buffers
- Basic Assay Protocol
- Assay Optimization
- Data Analysis
- Filtration Assays
- Non-Radiometric GTPγS Assays
- Literature References
Tissue Culture Assays
- Cell Culture Glossary
- Introduction to Cell Based Assays
- Requirements for Assay Proof of Concept for Cell Based Assays
- Tissue Culture Assay Development, Optimization and Validation Flow Chart
- Types of Tissue Culture Assays
- Batch Co-Transfection and Cell Banking Protocols
- Cryopreservation of Cells
- Cell Counting Protocols
- Factors for Cell Based Assay Development
- Typical Cell Culture Protocol
- Standard HTS Cell Culture Practices to Reduce Contamination Risk
- Example, Transient Transfection of Cells
- Transient Transfection Methods
- Transfection Cationic Matrix Reagents
- General Cell Biology Information
- References
Cell-Based Elisa (C-Elisa) and Westerns Blots for Quantitative Antigen Detection
- Introduction
- Cell Culture and Treatments
- Fixing and Staining
- Reagents
- Assay Validation
- Appendix 1. Immunoblot vs. Cytoblot Assays for Enzyme Inhibitor Potencies
- Appendix 2. Linearity and Sensitivity of Cell-Based Western Assays
- References
Additional Functional Assays for GPCRs and Ion Channels
- cAMP Detection
- β-Arrestin Recruitment
- IP-3/IP-1 Assays
- Impedance-Based Technologies
- Phospho-ERK Assays
Flipr™ Assays to Measure GPCR and Ion Channel Targets
- Overview: Flipr™ Assay Development
- Introduction
- Types of Flipr™ formats
- Reagents and Buffers for Method Development
- Method Development and Optimization
- Flipr™ Instrument Set-Up
- Potential Artifacts
- References
Immunoassay Methods
- Immunoassay Development, Optimization and Validation Flow Chart
- Introduction
- Immunoassay Parameters
- Reagents
- Instrumentation
- Immunoassay Formats
- Important Parameters for Development of an Immunoassay
- Initial Concept and Method Development of a Sandwich Immunoassay
- Results
- Second Experiment-Matrix Compatibility
- Initial Concept and Method Development of a Competitive Assay
- Development of a Competitive Immunoassay
- Method Validation (Pre-Study)
- Method Validation (In-Study)
- Pre-Study & In-Study Acceptance Criteria
- References
Guidelines for Cell Proliferation Assays
- Cell Proliferation Assays
- Methods for Evaluating Cell Proliferation
- Setting Up a Cell Proliferation Assay
Cell-Based RNAi Assays
- Introduction
- RNAi Loss-of-Function Screens
- RNAi Synthetic Lethality Screens
- Appendix
- Acknowledgements
- References
- Abbreviations
Data Standardization for Results Management
- Introduction
- Data Types and Associated Rules for Radioligand Binding Assays: Inhibition Mode
- Data Types and Associated Rules for Enzymatic Assays: Inhibition Mode
- Data Types and Associated Rules for In Vitro Functional Assays
- Glossary
Calculations and Instrumentation Used for Radioligand Binding Assays
- Introduction
- Radioactive Calculations
- Instrumentation
- Uniformity Plate
- Color Quench Correction
- Abbreviations
Assay Equipment and Instrumentation
- Introduction
- Common Equipment in HTS Labs
- Spectrophotometry
- pH meters
- Electronic Balances
- Microscopes
- References
In Vivo Assay Guidelines
Glossary of Quantitative Biology Terms
Sites Collaborating with NCGC
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Trial Sections- To be reviewed and approved by Editorial Board
Section 19 - Stem Cells, Regenerative Medicine & Chemical Biology -G. Sitta Sittampalam
