| Assay Method Information | |
| | In Vitro Kinase Assay |
| Description: | In vitro kinase assays were conducted using Kinase Profiler services provided by Eurofins. The experimental method was as follows: the small molecule compound to be tested (0.001-10 μM), the protein kinase to be tested were incubated with a buffer containing substrate, 10 mM magnesium acetate, and [γ-33P-ATP], the reaction was started by adding MgTPmix, after incubation for a period of time at room temperature, a 3% phosphate solution was added to the buffer to stop the reaction. Subsequently, 10 μL of the reaction mixture was quantitatively pipetted onto P30 filter paper, which was washed three times with 75 mM phosphate solution and then once with methanol. The P30 filter paper was air-dried and scintillation fluid was added for scintillation counting. The inhibitory activity of the compounds was expressed as the half maximal inhibitory concentration, IC50, which was fitted to the inhibition ratio corresponding to each concentration gradient. |
| Affinity data for this assay | |
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