Assay Method Information

Assay Name:  Biochemical Inhibition of Enzymatic Activities Assay
Description:  Table D: Corresponding biochemical inhibition of enzymatic activities of FLT3 (wt), FLT3 (D835Y), and FLT3 (ITD) were measured using recombinant protein constructs of kinase domains via activity based FLT3 kinase assay for compound screening and profiling via radiometric HotSpot™ kinase assay (Reaction Biology). Peptide substrate [EAIYAAPFAKKK]. Compounds were dissolved to 10 mM in DMSO. Compounds were tested in 10-dose IC50 mode with a 3-fold serial dilution starting at 0.3 μM. Control compound, Staurosporine, was tested in 10-dose IC50 mode with 4-fold serial dilution starting at 20 μM. Alternate control compounds were tested in 10-dose IC50 mode with 3-fold serial dilution starting at 20 μM. Reactions were carried out at 1 μM ATP.
Affinity data for this assay
 

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