Fluorescence Resonance Energy Transfer. Ad measure absorbance fluorescence luminescence htrf tr fret and alphascreen. The theory supporting energy transfer is based on the concept of treating an excited fluorophore as an.
We mutated an innocuous surface residue to cysteine and labeled separate pools with fluorescein donor and tetramethylrhodamine acceptor. Enzymatic cleavage of the scissile bond results in release of the fret quenching partner while the fluorescent product is retained on the cell surface allowing isolation of catalytically active clones by fluorescence activated cell sorting facs. The process can be summarized as.
The mechanism of fluorescence resonance energy transfer involves a donor fluorophore in an excited electronic state which may transfer its excitation energy to a nearby acceptor chromophore in a non radiative fashion through long range dipole dipole interactions.
Fluorescence resonance energy transfer. Therefore the excitation spectra of the acceptor molecule must overlap with the emission of. Bioluminescence resonance energy transfer bret is a valuable tool to detect protein protein interactions. As the efficiency of energy transfer is proportional to r 6 where r is the distance between donor.