Assay Method Information

Assay Name:  Kinase Assay
Description:  FLT3 kinase assay was obtained from Claiper Life Sciences (Catalog #200-0423). The assay was carried out with human recombinant FLT3 (0.287 nM), fluoresce in labeled peptide substrate (with a peptide sequence of EAIYAAPFAKKK, 1.5 μM) and test compounds (in serial dilution) using 384-well plates, quantified by Caliper technology. Kinase reaction was performed in 100 mM HEPES, pH 7.5, 4% DMSO, 0.003% Brij-35, 0.004% tween, 10 mM MgCl2, and 100 μM ATP (for IC50 determination), incubated at 28° C. for 90 minutes. After incubation, the reaction product was analyzed by electrophoretic mobility shift run on Caliper by manufacturer's protocol.
Affinity data for this assay
 

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