| Assay Method Information | |
| | Kinase Activity Assay |
| Description: | BRAF kinase assays were performed using the Z′-LYTE™ enzymatic assay (Invitrogen, USA). Briefly, kinase activity was monitored in a cascade system consisting of a mixture of inhibitor with BRAF or BRAFV600E/inactive MAP2K1 (MEK1)/inactive MAPK1 (ERK2)/Ser/Thr 03 peptide (Invitrogen) in 50 mM HEPES pH 7.5, 100 μM ATP, 10 mM MgCl2, 1 mM EGTA, 0.01% Brij-35. Titrations were performed using a 1:3 dilution. Assays were performed using Select Screen (Invitrogen). |
| Affinity data for this assay | |
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