Carla Jane Duval, Candy Laura Steffen, Karolina Pavic, Daniel Kwaku Abankwa
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Protocol to measure and analyze protein interactions in mammalian cells using bioluminescence resonance energy transfer.
Bioluminescence resonance energy transfer (BRET) allows to quantitate protein interactions in intact cells. Here, we present a protocol for measuring BRET due to transient interactions of oncogenic K-RasG12V in plasma membrane nanoclusters of HEK293-EBNA cells. We describe steps for seeding, transfecting, and replating cells. We then detail procedures for their preparation for BRET measurements on a CLARIOstar microplate reader and detailed data analysis. For complete details on the use and execution of this protocol, please refer to Steffen et al.1.
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