| Assay Method Information | |
| | Enzyme Activity Inhibitory Assay |
| Description: | Kinase Assay:Preparation of buffers, the buffer included 50 mM HEPES (pH 7.5), 0.01% Brij-35, 10 mM MgCl2, and 1 mM EGTA.After the buffer was formulated, the enzyme and substrate were mixed with different concentrations of the compound prepared in advance, and placed at room temperature for 15 minutes. ATP was added to start reaction, and the reaction solution was incubated at room temperature for 90 minutes (positive and negative controls were set). 10 μL of reaction system included 2.5 μL compound, 5 μL mixture of enzyme and substrate, and 2.5 μL ATP. After the reaction was completed, the antibody was added to test and incubated at room temperature for 60 min, then was detected by Evnvision and data was collected. Data analysis and mapping was conducted with XLfit5 software. |
| Affinity data for this assay | |
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