| Assay Method Information | |
| | Measurement of CSF1R Kinase Activity |
| Description: | Methods: 1) Kinase Reaction (10 μL System)2.5 μL of 4× test compound was added to each reaction well of the 384 plate, and the corresponding volume of 8% DMSO was added to the control well. The plate was placed on ice. 5 μL of a mixture of kinase/peptide substrate, 2.5 μL of kinase buffer and ATP solution were successively added to each well. Three control groups were set up: Group C1 contains only kinase buffer, group C2 contains the mixture of kinase/peptide substrate, kinase buffer and ATP, and group C3 contains 5 μL of PP solution. After adding the reaction components, the 384-well plate was sealed and incubated at 25-30° C. for 1 h in dark.2) Development Reaction5 μL of Development solution was added to each well, sealed and incubated at 25-30° C. for another 1 h in dark.3) Reaction Stopping and Plate Reading5 μL of stopping solution was added to each well. The Coumarin value (excitation at 400 nm, emission at 445 nm) and Fluorescein value (excitation at 400 nm, emission at 520 nm) were measured, respectively. |
| Affinity data for this assay | |
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