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Kinase binding — HitHunter EFC assays

Enzyme Fragment Complementation for kinase binding

Kinase Binding EFC Chemiluminescence HitHunter Assays LCK p38 MAPK pKC alpha Src / c-

HitHunter Kinase Binding Assays are used to identify and characterize compounds which bind to a kinase-binding pocket. Inhibitor probes are conjugated to the ED fragment of E. coli b-galactosidase (b-gal), which then competes with free compound for ATP-binding sites on the kinase of interest.

The HitHunter 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.



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 and c- Kit

Select another in vitro kinase assay