| Assay Method Information | |
| | Biochemical Kinase Assay |
| Description: | Specific Assay Method:Compound formulation: The test compounds were dissolved in DMSO to prepare a 20 mM stock solution. Before use, the compounds were diluted in DMSO to 0.1 mM, and diluted into 11 concentrations with a 3-fold gradient. When addition, diluentions of 4-fold final concentration were then made with buffer solution.Kinase assay method: After preparing the buffer, the enzyme was mixed with pre-formulated and diluted compounds of different concentrations in duplicate for each concentration, and placed at room temperature for 30 minutes. The corresponding substrate and ATP were added thereto, and reacted at room temperature for 60 minutes (both a negative and a positive control were set). After the reaction, antibody was added for detection, Envision detection was carried out after 60 minutes incubation at room temperature, and data was collected. Data analysis and fitting were made according to XLfit5 software, wherein, A represents IC50≤5 nM, B represents IC50 is 5-50 nM, C represents IC50 is 50-100 nM, D represents IC50>100 nM. |
| Affinity data for this assay | |
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