| Assay Method Information | |
| | CDK7/CyclinH/MAT1 Enzyme Reaction System |
| Description: | Enzymes, substrates (MBP), 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 CDK7/CyclinH/MAT1 enzyme (20 ng), 2 μL mixture of substrate and ATP (10 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 120 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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