HitHunter™ Kinase Binding Assay

Functional kinase assays are based on detection of substrate phosphorylation by the enzyme. However, when substrate phosphorylation is not known or when mutant kinases or unactive kinases are utilized, activity based assays cannot be used. DiscoveRx offers a novel binding-based method that directly measures the binding potency of compounds to the ATP binding site of the kinase through competition with a conjugated binding probe enabling you to include unactive and low activity kinases in your screening campaigns.

Product Features

•Measure competitive inhibitor binding to the kinase active site
•Homogeneous assay
•No antibody or substrate required
•Chemiluminescence, gain-of-signal readout
•Sensitive assay
•Scalable assay format

Applications:

•Screen unactive kinases
•Compare compound potency between active and unactive forms of the kinase
•Also ideal for non-ATP pocket binders

How does the assay work?

The Kinase Binding Assay Platform utilizes EFC technology to homogeneously measure binding of inhibitors to the ATP binding site. In the assay, a competitive binding reaction is set-up with a compound, an enzyme donor (ED)-ligand conjugate, and a kinase. Incubation allows competition between the free ligand and ED-ligand conjugate for the ATP binding pocket of the kinase. Enzyme acceptor (EA) is added with chemilumescent substrate. Complementation of EA with the displaced ED-ligand conjugate in the presence of chemilumescent substrate generates signal, which can be measured using any standard luminometer.

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Available Collaterals

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