Hello, I am measuring heterodimerization of GPCRs by FRET acceptor photobleaching, using the CFP/YFP pair.

I am using cells transfected only with GPCR-CFP as negative control, in order to measure CFP indirect increase of fluorescence after YFP bleaching. This negative control works fine, producing negative values of FRET efficiency.

I am wondering if 4.5% of FRET efficiency that I can measure between GPCR1-CFP and GPCR2-YFP is too low to be considered a positive value. Indeed, I can see in the literature that positive values of FRET start from 10%.

thank you !!

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