| Assay Method Information | |
| | Enzyme Activity Test |
| Description: | 1) The compound solution and the positive control were diluted 8.3 times with double distilled water and then added to the 384 well plate, each concentration was 2 uL/well; (2) 6 nM CDK7/Cyclin H/MAT1 kinase solution was added to the compound well and the positive control well respectively; (3) incubating at room temperature for 0 and 60 minutes; (4) 2 mM ATP and 2 pM 5-FAM-CDK7 peptide substrate (5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK) solution (8 nM CDK9/Cyclin Tl polymer and 2 pM 5-FAM GSRTPMY-NH2 peptide substrate were used in CDK9 inhibition reaction; 50 nM CDK12 (aa686-1082)/Cyclin K polymer and 2 pM 5-FAM GSRTPMY-NH2 peptide substrate were used in CDK12 inhibition reaction; 0.5 nM CDK2/Cyclin El polymer and 2 pM 5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK peptide substrate were used in CDK2 inhibition reaction) were added; (5) Incubating the 384 well plate at 25° C. for 30 minutes; (6) 4 uL 120 mM EDTA was added to stop the kinase reaction; (7) the conversion rate was read with Caliper/LabChip EZ Reader (Perkin Elmer); (8) IC50 value was obtained by curve fitting using GraphPad Prism 8 software. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |