| Assay Method Information | |
| | Enzymatic Inhibitory Activity (IC50) Assay |
| Description: | The operation method is briefly described as follows:The powders of the compound were dissolved in 100% DMSO to make a 10 mM stock solution. The initial test concentration of the compound was 10,000 nM, 3-fold serial dilution, 10 concentrations, and repeated hole detection. The serially diluted compounds and kinases were mixed in the Optiplate-384F well plate and incubated at room temperature for 10 minutes, then ATP and Kinase substrate30 (GL Biochem, Cat.117885) mixture were added, mixed homogeneously and then reacted at room temperature for 20 minutes. A stop detection solution was added to stop the enzymatic reaction, and the conversion rate was read with Caliper EZ Reader II.Data analysis: %Inhibition=Conversion %_max-Conversion %_sampleConversion %_max-Conversion %_min 100[0301]wherein: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min: the average value of the negative control wells, representing the conversion rate readings of the wells without enzyme activity; Conversion%_max: the average ratio of the positive control wells, representing the conversion rate readings of the wells without compound inhibition. |
| Affinity data for this assay | |
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