| Assay Method Information | |
| | KIT Tyrosine Kinase Assay |
| Description: | Recombinant KIT protein was diluted to 1 μM with Assay Buffer. Poly (Glu4-Tyr) peptide and ATP were diluted to 300 microgram/mL and 250 μM, respectively, with Assay Buffer, then mixed to make the substrate solution. In a 384-well plate, 2 μL of working compounds and 5 μL of recombinant KIT protein were mixed and incubated for 30 minutes at room temperature. After the preincubation, 3 μL of substrate/ATP mixture was added and incubated for 1 hour at room temperature. The tyrosine phosphorylation by KIT kinase was quantified by ADP-Glo kinase assay kit following the manufacturer's protocol. Briefly, the phosphorylation reaction was terminated by adding 10 μL ADP-Glo reagent and incubated for 60 minutes at room temperature. 20 μL of Kinase Detection Reagent was added to each well and luminescence was measured by plate reader. The assay control for this experiment was 2.5% DMSO and Assay Buffer was used as the blank. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |