| Assay Method Information | |
| | Inhibitory Assay |
| Description: | An inhibitory activity of the above compound in-vitro against Aurora A kinase activity was measured with reference to a method described in JP-A-2008-81492. In measuring inhibitory activity of the compound, first, the test compound was serially diluted by dimethylsulfoxide (DMSO). Then, to a reaction buffer [50 mM Tris-hydrochloric acid buffer (pH 7.4), 15 mM magnesium acetate, 0.2 mM ethylenediamine-N,N,N',N'-tetraacetic acids (EDTA)], purified human Aurora A protein, FL-Peptide 21 (Caliper Life Sciences, Inc., a final concentration: 100 nM), ATP (a final concentration: 5 uM) and the DMSO solution (a final concentration of DMSO: 5%) of the test compound were added. The mixture was incubated at 25 C. for 50 minutes to perform a kinase reaction. To the mixture, IMAP (registered trade mark) Progressive Binding Reagent diluted 500x with IMAP (registered trade mark) Progressive Binding Buffer A (manufactured by Molecular Devices) was added to terminate the kinase reaction. |
| Affinity data for this assay | |
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