| Assay Method Information | |
| | Test of AKT Kinase Inhibiting Activity |
| Description: | After all the reagents were prepared according to the above method, except for the enzyme, the reagents were equilibrated to the room temperature and loaded. a) first, a compound stock solution (10 mM DMSO solution) was diluted with DMSO to obtain a 100 µM compound solution, the compound solution was diluted with the 1× kinase reaction buffer to obtain a 2.5 µM compound working solution (containing 2.5% DMSO). A 2.5% DMSO solution was prepared from the 1× kinase reaction buffer, and the 2.5 µM compound working solution was diluted 7 times with the 2.5% DMSO solution according to a 4-fold gradient to obtain compound working solutions at 8 concentrations (2500 nM, 625 nM, 156 nM, 39 nM, 9.8 nM, 2.4 nM, 0.6 nM, and 0.15 nM). Except for control wells, 4 µL of diluted compound working solution was placed in each reaction well, and 4 µL of previously prepared 2.5% DMSO/kinase buffer was placed in each control well. |
| Affinity data for this assay | |
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