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How to cite this document:
Assay Guidance Manual Version 5.0, 2008, Eli Lilly and Company and NIH Chemical Genomics Center.
Available online at: http://www.ncgc.nih.gov/guidance/manual_toc.html (last accessed [insert date here])
- INTRODUCTION
- 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
- DETERMINATION OF EC50/IC50
- PRODUCTION MONITORING
- ENZYME ASSAY DEVELOPMENT FLOW CHART
- INTRODUCTION
- CONCEPT
- REAGENTS AND METHOD DEVELOPMENT
- DETECTION SYSTEM LINEARITY
- ENZYME REACTION PROGRESS CURVE
- MEASUREMENT OF KM AND VMAX
- DETERMINATION OF IC50 FOR INHIBITORS
- INHIBITION CONSTANT (KI)
- IC50 DETERMINATION FOR SAR
- OPTIMIZATION EXPERIMENTS
- ASSAY VALIDATION
- 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
- STEPS TO ASSAY DEVELOPMENT
- INTRODUCTION
- MATERIALS AND REAGENTS
- MEMBRANE PREPARATIONS AND ASSAY BUFFERS
- BASIC ASSAY PROTOCOL
- ASSAY OPTIMIZATION
- DATA ANALYSIS
- FILTRATION ASSAYS
- NON-RADIOMETRIC GTPγS ASSAYS
- LITERATURE REFERENCES
- 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
- 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
- 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 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
- 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
- INTRODUCTION
- DATA TYPES AND ASSOCIATED RULES FOR RADIOLIGAND BINDING ASSAYS: INHIBITION MODE
- ATA TYPES AND ASSOCIATED RULES FOR ENZYMATIC ASSAYS: INHIBITION MODE
- DATA TYPES AND ASSOCIATED RULES FOR IN VITRO FUNCTIONAL ASSAYS
- GLOSSARY
- GLOSSARY OF MOA TERMS
- OVERVIEW OF MOA IN DRUG DISCOVERY
- TYPES OF INHIBITION
- PERFORMING MOA STUDIES
- REFERENCES
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