HitHunter™ EFC Kinase Assays by DiscoveRx Corporation
Manufacturer DiscoveRx Corporation
Overview & 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™ EFC Assays for functional kinase activity are an antibody-based competitive binding format that uses EFC chemiluminescent detection. EFC detection provides sensitivity throug... Read more
The supplier does not provide quotations for this product through SelectScience. Browse these related products instead
Related Products
.jpg.ashx?width=100&height=100&bgcolor=white)
HitHunter™ EFC Kinase Assays
Overview & 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™ EFC Assays for functional kinase activity are an antibody-based competitive binding format that uses EFC chemiluminescent detection. EFC detection provides sensitivity through signal amplification. The EFC signal generated is proportional to the amount of phosphopeptide generated by the kinase enzyme.
Features and Benefits
» Proprietary Antibody/substrate pairs with the broadest coverage of the kinome
» Minimize interference
• Chemiluminescent signal eliminates optical interference due to fluorescent compounds with no artificial signal due to non-specific binding of beads or a secondary label
» Homogeneous
• Add and mix protocols simplify automation for quicker assays without the need for washing, centrifugation or filtering.
» Flexible
• Scaleable in 96-, 384-, and 1536-well microplate formats
» Robust reproducible signal
• EFC signal is amplified enzymatically
• producing large signal to background ratios and Z’> 0.7
» Convenient
• Use existing liquid handling either in batch or HTS mode
• with a wide range of luminometers, mutli-label microplate readers and
• CCD cameras.
Assay Design
HitHunter Kinase EFC 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 40 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 EFC product inserts list the limits of common additives, solvents and concentrations that do not affect assay performance.
Product Details
Kit Contents
» Phosphopeptide ED conjugate, EFC Label
» Antibody
» Kinase substrate
» EFC detection reagents
» Phosphopeptide Standard
Instrument Requirements
» Luminometer, Multimode Microplate Reader or CCD Camera
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™ EFC Assays for functional kinase activity are an antibody-based competitive binding format that uses EFC chemiluminescent detection. EFC detection provides sensitivity through signal amplification. The EFC signal generated is proportional to the amount of phosphopeptide generated by the kinase enzyme.
Features and Benefits
» Proprietary Antibody/substrate pairs with the broadest coverage of the kinome
» Minimize interference
• Chemiluminescent signal eliminates optical interference due to fluorescent compounds with no artificial signal due to non-specific binding of beads or a secondary label
» Homogeneous
• Add and mix protocols simplify automation for quicker assays without the need for washing, centrifugation or filtering.
» Flexible
• Scaleable in 96-, 384-, and 1536-well microplate formats
» Robust reproducible signal
• EFC signal is amplified enzymatically
• producing large signal to background ratios and Z’> 0.7
» Convenient
• Use existing liquid handling either in batch or HTS mode
• with a wide range of luminometers, mutli-label microplate readers and
• CCD cameras.
Assay Design
HitHunter Kinase EFC 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 40 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 EFC product inserts list the limits of common additives, solvents and concentrations that do not affect assay performance.
Product Details
Kit Contents
» Phosphopeptide ED conjugate, EFC Label
» Antibody
» Kinase substrate
» EFC detection reagents
» Phosphopeptide Standard
Instrument Requirements
» Luminometer, Multimode Microplate Reader or CCD Camera
Product Overview
