Utilizing NanoBRET technology to measure BRAF autoinhibition in live cells

BRAF is frequently activated via mutation in human cancer and the RASopathy syndromes; however, for BRAF activation to occur, autoinhibitory interactions between the regulatory and catalytic domains must be relieved. We have generated isolated BRAF catalytic and regulatory domain proteins where the catalytic domain is tagged with a NanoLuc energy donor and the regulatory domain is fused to a HaloTag for labeling with an energy acceptor. These constructs can be used in a proximity-based, NanoBRET assay for measuring BRAF autoinhibition in live cells. This data set contains raw scans of blots that show protein expression levels in control experiments for assay optimization.