| Assay Method Information | |
| | In Vitro Kinase Detection Assay |
| Description: | Kinases BTK wt (Carna, Cat. No 08-180) and BTK C481S (Carna, Cat. No 08-547) were prepared into a 2.5× kinase solution, and substrates FAM-P2 (GL Biochem, Cat. No. 112394) and ATP ((Sigma, Cat. No. A7699-1G) were prepared into a 2.5× substrate solution, respectively. 5 μL of compounds at different concentrations were added to a 384-well plate. 10 μL of 2.5× kinase solution was added, and the resulting mixture was incubated at room temperature for 10 min. 10 μL of 2.5× substrate solution was added, and the mixture was incubated at 28° C. for an appropriate period of time. The reaction was stopped by adding 30 μL of stop buffer, and the detection was carried out by using Caliper EZ reader2. The IC50 value was calculated by using XLFit excel add-in version 5.4.0.8 software. |
| Affinity data for this assay | |
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