BRAF (V600E) NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview
Target
BRAF (V600E)
HGNC Symbol
BRAF (V600E)
Synonyms
B-Raf; BRAF1; RAFB1
Cell Line
HEK293 human epithelial cell line from ATCC
Assay Properties
Assay Format
384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay
Assay Protocol
HEK293 cells transiently expressing NanoLuc-BRAF(V600E) Fusion Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-10 and then treated with reference compound PD-173955 for 1 hour. The BRET signal was measured on an Envision 2104 Multilabel Reader. IC50 value was calculated and IC50 curve was plotted using the GraphPad Prism 4 program based on a sigmoidal dose response equation.
Readout
Binding of compound to target kinase
Reference Compound IC50s
| Compounds | IC50 (M) |
|---|---|
| PD-173955 | 1.359e-008 |
Screening Location
Malvern, USA
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