Figure 2. c-Kit kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl 2 , 3 mM MnCl 2 , 3 µM Na-orthovanadate, 1.2 mM DTT, ATP (variable), 2.5 µg/50 µl PEG20.000, Substrate: Tetra (LRRWSLG), 5 µg/50 µl, recombinant c-Kit: 200 ng/50 µl.
Figure 1. Dose dependence curve of c-Kit kinase activity: DELFIA® data generated using phospho-Tyrosine mAb (p-Tyr-100) #9411 to detect phosphorylation of substrate peptide #1364 by c-Kit kinase. In a 50ul reaction, increasing amounts of c-Kit and 1.5uM substrate peptide were used per reaction at room temperature for 30 minutes (DELFIA® is a registered trademark of PerkinElmer, Inc.)