| Assay Method Information | |
| | B-RAF (V600E) Kinase Inhibition Assay |
| Description: | The biological activities (IC50s) of the compounds were determined against the B-RAF (V600E) mutant enzyme in vitro. GST-MEK1, B-RAF (V600E), and inhibitor were added to the wells of glutathione-coated plate, and the plate was preincubated for 10 min. ATP was added to each well, and the plates were incubated for 45 min. Primary antibody (rabbit anti-phospho MEK1/2) and Eu-labeled anti-rabbit secondary antibody were added to the plates. After incubation, DELFIA enhancement solution (Perkin-Elmer, Turku, Finland) was added. The europium counts were measured on a Victor 2 reader (Perkin-Elmer, Turku, Finland). |
| Affinity data for this assay | |
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