| Assay Method Information | |
| | CK2a2 (CSNK2a2) Enzyme Assay |
| Description: | Briefly, CK2a2 (CSNK2a2) (7 nM) in the enzymatic buffer solutions (40 mM Tris, pH 7.5, 20 mM MgCl2, 0.1 mg/ml BSA, 50 μM DTT) were mixed with various concentrations of inhibitors (dissolved in 100% DMSO). These solutions were incubated for 30 minutes at 25° C., and subsequently a mixture of substrate peptide and ATP was added with the final peptide and ATP concentrations at 0.04 mg/ml and 15 μM, respectively. The final reaction mixture of enzyme-substrate-ATP-compound was incubated for 90 minutes at 25° C.Afterwards ADP-Glo™ Reagent was added and the mixture was incubated at 25° C. for 60 minutes. Then Kinase Detection Reagent was added, and the final mixture was incubated at 25° C. for 60 minutes. The luminescence signal of the final solution was measured on an Envision instrument (PerkinElmer). The percent (%) inhibition at each concentration of a compound is calculated relative to the luminescence signal in the Max and Min control wells contained within each assay plate. The Max control wells contain both enzyme and substrate as 0% inhibition, and the Min control wells only contain substrate without enzyme as 100% inhibition. |
| Affinity data for this assay | |
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