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A general Bioluminescence Resonance Energy Transfer (BRET) protocol to measure and analyze protein interactions in mammalian cells

Duval, C. J.; Steffen, C. L.; Pavic, K.; Abankwa, D. K.

2024-07-05 cell biology
10.1101/2024.07.05.602189 bioRxiv
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Bioluminescence resonance energy transfer (BRET) allows to quantitate protein interactions in intact cells. Here we provide a step-by-step protocol for measuring BRET due to transient interactions of oncogenic K-RasG12V in plasma membrane nanoclusters of HEK293-EBNA cells. We describe how to seed, transfect and replate cells, followed by their preparation for BRET-measurements on a microplate reader and detailed data analysis steps. For details on how to apply this protocol, please refer to Steffen et al., 2024 1. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/602189v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@152e8c2org.highwire.dtl.DTLVardef@2f2aacorg.highwire.dtl.DTLVardef@9a8e9borg.highwire.dtl.DTLVardef@108833f_HPS_FORMAT_FIGEXP M_FIG C_FIG

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