FRET

FRET - Fluorescence Resonance Energy Transfer

• method for assessment of protein-protein interaction using fluorescent tags, eg. CFP and YFP;
• FRET proves if molecules are within 10nm or closer e.g. heterodimers, DNA bound molecules;
• accuracy of visual co-localisation with light microscopy is ~500nm, so molecules may not really be together;

What probes are required for cytometry based FRET?

2 probes:

• Donor molecule, which is excited
• Acceptor molecule, which emits

Donor emission spectrum has to overlap acceptor excitation spectrum.

Energy passed on in a non-radiative manner.