| Assay Method Information | |
| | Inhibitory Activity Assay |
| Description: | 1. Methods(1) 4 μL of prepared kinase buffer solution or 4 μL of kinase solution (100% inhibition control) were added into a 384-hole plate; 2 μL of the compound or 2 μL of buffer without the compound (0% inhibition control) was added into the holes; 2 repetitive holes were set for all the samples or control above.(2) Incubated at 25° C. for 5 min;(3) 2 μL of ATP/substrate/MgCl2/MnCl2/SEB/DTT mixed solution were added into the holes;(4) Centrifugated at 1000 rpm for 1 min, and incubated by vibration at 30° C. for 30 min;(5) 8 μL of XL-665/antibody mixed solution were added into the holes;(6) Incubated at 25° C. for 1 h;(7) The signals at 665 nm and 620 nm were read on PHERAstar FS;(8) The data was analyzed according to the instructions of the kit, and the fitting calculation of IC50 was performed using GraphPad Prism5. |
| Affinity data for this assay | |
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