- TR-FRET = Time Resolved – Fluorescence Resonance Energy Transfer.
- 384 well competition assay.
- Anti-6xHis antibody, labelled with a fluorescent donor, F(d), binds the tagged enzyme. The enzyme ligand is tagged with a fluorescent acceptor, F(a). Complex formation results in energy transfer from donor to acceptor fluorophore non-radiatively. The ratio between light emission (wavelength B/A) is calculated. Kd value of the inhibitor is obtained.
Click here to see how this compares with the functional PPIase assay.
Schematic of the TR-FRET Assay