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FRET-based translocation assay principle. Eukaryotic cells loaded with the fluorescent substrate CCF2-AM display a green fluorescence (emission at 530 nm) when excited at 405 nm. Upon translocation of a β-lactamase–based chimeric protein into the host cell cytoplasm, the cephalosporin core of CCF2-AM is cleaved, resulting in blue fluorescence (emission at 447 nm) upon the same excitation (11).
