| Assay Method Information | |
| | In Vitro Kinase Assay |
| Description: | Inhibition of kinase activity against a variety of recombinant kinases [FLT3, FLT3 D835Y, anaplastic lymphoma kinase (ALK), insulin receptor, and epidermal growthfactor receptor (EGFR)] was measured using homogeneous time-resolved fluorescence (HTRF) assays [Choi et al., Bioorg. Med. Chem. Lett., 20:2033-2037]. Assays consist of enzymes mixed with serially diluted compounds and peptide substrates in a kinase reaction buffer (250 mM HEPES (pH 7.0), 0.5 mM orthovanadate, 0.05% bovine serum albumin, 0.1% NaN3). Following the addition of reagents for detection, the TR-FRET signal was measured using an EnVision multi-label reader (Perkin Elmer, Waltham, MA). |
| Affinity data for this assay | |
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