Assay Method Information | |
| CDK2/CyclinA2 Enzyme Reaction System |
Description: | Enzymes, substrates, ATP, and inhibitors were diluted by kinase buffer in the kit. The compounds to be tested were diluted 5-fold with a multi-channel pipette to the eighth concentration, that is, from 50 μM to 0.65 nM, the concentration of DMSO was 5%, and a double-hole experiment was set up. 1 μL of each concentration gradient of inhibitors, 2 μL of CDK2/CyclinA2 enzyme (1.6 ng), 2 μL mixture of substrate and ATP (50 μM ATP, 0.1 μg/μL substrate) were added to the microplates, and the final concentration gradient of the compound was 10 μM to 0.13 nM. The reaction system was reacted at 25° C. for 60 minutes. After the reaction, 5 μL of ADP-Glo reagent was added to each well, the reaction was carried out at 25° C. for 40 minutes, after the reaction was completed, 10 μL of kinase detection reagent was added to each well, the reaction was carried out at 25° C. for 30 minutes, the multiple label analyzer was used to read the chemiluminescence with an integration time of 0.5 seconds |
Affinity data for this assay | |
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