HitHunter™ Kinase Binding Assays for Unactive Kinases

HitHunter™ Kinase Binding Assays for Unactive Kinases

DiscoveRx Corporation

Description and Overview When little or no information is known about the kinase substrate and its associated antibody, an alternative approach is inhibitor binding directly or indirectly to the ki...read more

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Description

 

Description and Overview
When little or no information is known about the kinase substrate and its associated antibody, an alternative approach is inhibitor binding directly or indirectly to the kinase. Currently there is a growing need to screen for inhibitors that bind to unactive and low activity kinases due to the potential for augmented kinase inhibitor selectivity at pathologically important mutant forms of the enzyme. The HitHunter EFC platform has been extended using synthetic probes that bind to the ATP binding pocket of inactive, active, or unactive kinases.

Currently, 4 different ED-inhibitor probes are available in an assay format:
ED-SB202190 for p38 MAPK
ED-Staurosporine for PKCalpha and GSK3alpha
ED-Pyrazoloprimidine for LCK
ED-PD166285 for Src, c-Kit, and Abl 

Profiling and custom labeling services are also available upon request.

Features and Benefits
» Measure competitive inhibitor binding to the kinase active site
» Homogeneous assay
• No antibody or substrate required
» Compatible with active, unactive and inactive kinases
» Chemiluminescence, gain-of-signal readout
• No fluorescence interference
• Sensitive assay
» Scalable assay format

Kinase Binding using HitHunter EFC technology
The Kinase Binding Assay Platform utilizes EFC technology to homogeneously measure binding of inhibitors to the ATP binding site . In the assay, the ED-inhibitor conjugate and your compounds compete for binding to the ATP binding site. If the conjugate is not bound to the kinase, it is free to recombine with EA (inactive EFC detection enzyme), resulting in an active ß-gal enzyme, which hydrolyzes the luminescent substrate. The signal produced by active EFC enzyme is proportional to the amount of compound bound to your kinase.

Kit Contents
» ED inhibitor probe, EFC Label
» EFC detection reagents
» Assay Buffer

Instrument Requirements
» Luminometer, Multimode Microplate Reader or CCD Camera

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