Bioluminescence Resonance Energy Transfer (BRET)-Based Assay for Measuring Interactions of CRAF with 14-3-3 Proteins in Live CellsRussell Spencer-Smith 1,2,3
1Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, 2Hollings Cancer Center, Medical University of South Carolina, 3Laboratory of Cell and Developmental Signaling, Center for Cancer Research, National Cancer Institute-Frederick
This protocol describes a BRET-based assay for measuring the interactions of the CRAF kinase with 14-3-3 proteins in live cells. The protocol outlines steps for preparing the cells, reading BRET emissions, and data analysis. An example result with identification of appropriate controls and troubleshooting for assay optimization is also presented.
