| Assay Method Information | |
| | BRAF-Inhibiting Activity Assay |
| Description: | The BRAF-inhibiting activity of the compounds listed in Table 6 below was evaluated by the time-resolved fluorescence-fluorescence resonance energy transfer assay as described below. The test compound, BRAF (Eurofins Genomics KK.) and MEK1 (Thermo Fisher Scientific Inc.) were mixed in ATP-containing buffer and reacted for 90 minutes at 30° C. LANCE (registered trademark) Eu-Phospho-MEK1/2(Ser217/221) antibody (Perkin-Elmer) was then added, and reaction was continued for 60 minutes at room temperature. Following the reaction, the fluorescence resonance energy transfer was measured with a fluorescent plate reader and the 50% inhibition concentration (IC50) was calculated based on the percent inhibition relative to a test compound-free control. |
| Affinity data for this assay | |
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