HitHunter™ FP Kinase Assays by DiscoveRx Corporation
Manufacturer DiscoveRx Corporation
Overview and Description Kinase enzymes are known to have broad substrate specificity and can phosphorylate a number of non-physiological substrates in vitro. When evaluating kinase targets, analysis is often hindered by the lack of suitable antibody/ kinase substrate pairs in a homogeneous, non-radioactive format. HitHunter™ Kinase FP Assays for functional kinase activity are an antibody-based competitive binding format using Fluorescence Polarization (FP) detection. These assays are developed ... Read more
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HitHunter™ FP Kinase Assays
Overview and Description
Kinase enzymes are known to have broad substrate specificity and can phosphorylate a number of non-physiological substrates in vitro. When evaluating kinase targets, analysis is often hindered by the lack of suitable antibody/ kinase substrate pairs in a homogeneous, non-radioactive format.
HitHunter™ Kinase FP Assays for functional kinase activity are an antibody-based competitive binding format using Fluorescence Polarization (FP) detection. These assays are developed using fluorescein tracers with 99% purity and a high affinity for the corresponding phosphopeptide.
Features and Benefits
» Proprietary Antibody/substrate pairs with the broadest coverage of the kinome
» Measure Enzyme Kinetics
» Homogeneous
• Simple reagent additions, mixing and measurement for HTS friendly formats
» Scaleable
• Automation friendly protocols that are scaleable in
• 96-, 384-, and 1536-well formats
» Minimize compound interference
• Red-shifted tracers minimize interference from fluorescent compounds
Assay Design
HitHunter Kinase FP Assays are easy to run using simple reagent additions. After the kinase reaction, there are two reagent additions for detection.
Assay Parameters
Kinase Reaction Volume 20 µL
Total Assay Volume 50 µL
[kinase enzyme] range varies by enzyme prep
[ATP] range < 50 –100 µM
EFC Assay time ~a minimum of 1 hour after the kinase reaction
Kinase Reaction
The kinase reaction is composed of kinase substrate, enzyme, ATP, compound and any co-factor required for activating phosphorylation. Because the components of the kinase reaction are specific to each kinase, each component and incubation time for the kinase should be optimized. HitHunter FP product inserts list the limits of common additives, solvents and concentrations that do not affect assay performance.
Product Details
Kit Contents
» Phosphopeptide Tracer
» Antibody
» Matched substrate
» Phosphopeptide Standard
Instrument Requirements
» Fluorescence Polarization Detection
Excitation: 485 nM
Emission: 530 nM
Kinase enzymes are known to have broad substrate specificity and can phosphorylate a number of non-physiological substrates in vitro. When evaluating kinase targets, analysis is often hindered by the lack of suitable antibody/ kinase substrate pairs in a homogeneous, non-radioactive format.
HitHunter™ Kinase FP Assays for functional kinase activity are an antibody-based competitive binding format using Fluorescence Polarization (FP) detection. These assays are developed using fluorescein tracers with 99% purity and a high affinity for the corresponding phosphopeptide.
Features and Benefits
» Proprietary Antibody/substrate pairs with the broadest coverage of the kinome
» Measure Enzyme Kinetics
» Homogeneous
• Simple reagent additions, mixing and measurement for HTS friendly formats
» Scaleable
• Automation friendly protocols that are scaleable in
• 96-, 384-, and 1536-well formats
» Minimize compound interference
• Red-shifted tracers minimize interference from fluorescent compounds
Assay Design
HitHunter Kinase FP Assays are easy to run using simple reagent additions. After the kinase reaction, there are two reagent additions for detection.
Assay Parameters
Kinase Reaction Volume 20 µL
Total Assay Volume 50 µL
[kinase enzyme] range varies by enzyme prep
[ATP] range < 50 –100 µM
EFC Assay time ~a minimum of 1 hour after the kinase reaction
Kinase Reaction
The kinase reaction is composed of kinase substrate, enzyme, ATP, compound and any co-factor required for activating phosphorylation. Because the components of the kinase reaction are specific to each kinase, each component and incubation time for the kinase should be optimized. HitHunter FP product inserts list the limits of common additives, solvents and concentrations that do not affect assay performance.
Product Details
Kit Contents
» Phosphopeptide Tracer
» Antibody
» Matched substrate
» Phosphopeptide Standard
Instrument Requirements
» Fluorescence Polarization Detection
Excitation: 485 nM
Emission: 530 nM
Product Overview
