| Assay Method Information | |
| | c-Met Z-LYTE Assay |
| Description: | The c-Met kinase activity of five target compounds and three positive compounds were evaluated using standard Z-LYTE Assays (fluorescence resonance energy transfer, FRET). The test compounds or reference controls were dissolved indimethyl sulfoxide (DMSO) as stock solution. The DMSO solution was serial diluted (duplicated with 3-fold dilution) to make that the concentrations were from 10 然 to 0.17 nM in 11 doses. The enzyme reaction mixture of c-Met Z-LYTE assay contained 2.5 nM c-Met, 1 然 Peptide substrate (Tyr 6) and 17 然 ATP in a buffer containing 50 mM HEPES (pH 7.0), 10 mM MgCl2, 0.01% Brij-35, 1 mM EGTA at a final volume of 10 無. The enzyme reaction were carried out at room temperature in black Proxiplate 384-Plus plate (PerkinElmer) for 90 min. 5 無 of the detectionreagents A (Invitrogen) was added at final concentrations of 64-fold dilution. The plates were incubated at room temperature for 60 min and then read in the Envision plate reader (PerkinElmer). |
| Affinity data for this assay | |
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