Application Note: LanthaScreen™ TR-FRET Assay From Invitrogen On Synergy 4

By Paul Held Ph. D., Senior Scientist, Applications Dept., BioTek Instruments, Inc.

Here we describe the use of the Synergy™ 4 Hybrid Microplate Reader to measure the output from Invitrogen's LanthaScreen™ TR-FRET assay technology. LanthaScreen™ technology uses time-resolved fluorescence resonance energy transfer (TR-FRET) to study multiple target classes including protein kinases, nuclear hormone receptors, as well as proteases and ubiquitinated proteins.

LanthaScreen™ TR-FRET works on the principles that when suitable pairs of fluorophores are in close proximity of one another, excitation of the terbium chelate donor fluorophore results in energy transfer to the fluorescein or GFP acceptor fluorophore. The benefit of this assay is the long fluorescence lifetime of the terbium lanthanide donor fluor excited at 340 nm. This allows measurement of the signal long after the background fluorescence has dissipated. The time-resolved measurement reduces assay interference (e.g. fluorescent compounds) and increases data quality.