A flow cytometric method to detect protein‐protein interaction in living cells by directly visualizing donor fluorophore quenching during CFP→YFP fluorescence resonance energy transfer (FRET)

Abstract

We have established a novel flow cytometric approach to assess donor CFP quenching during CFP-->YFP FRET, which can be used for the calculation of FRET efficiency and relative biological molecular distance between CFP and YFP moieties. This method can be used not only to analyze cells that express a CFP and YFP fusion protein, but also independent CFP-coupled and YFP-coupled interacting proteins.