| Assay Method Information | |
| | In Vitro Activity Assay |
| Description: | The kinase buffer in the kit was used to dilute the enzyme, substrate, ATP and inhibitor.The compound to be tested was diluted 5-fold to the eighth concentration with a discharge gun, i.e., from 50 μM to 0.65 nM, with a final DMSO concentration of 5%, and double duplicate wells experiment was set. 1 μL of each concentration gradient of the inhibitor, 2 μL of c-Met enzyme (4 ng), 2 μL of a mixture of the substrate and ATP (10 μM ATP, 0.2 μg/μL poly E4Y1) were added to the microplate, and the final concentration gradient of the compound of 10 μM was diluted to 0.13 nM. The reaction system was placed at 30° C. and the reaction was carried out for 60 minutes. When the reaction was completed, 5 μL of ADP-Glo reagent was added to each well and the reaction was carried out at 30° C. for another 40 minutes, when the reaction was completed, 10 μL of kinase detection reagent was added to each well, then the reaction was carried out at 30° C. for still another 30 minutes, chemiluminescence was read by the PerkinElmer Envision multi-label analyzer with an integration time of 0.5 seconds. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |