Improving lanthanide-based resonance energy transfer detection by increasing donor-acceptor distances.
Lanthanide-based resonance energy transfer (LRET) is an established method for measuring or detecting proximity between a luminescent lanthanide (energy donor) and an organic fluorophore (energy acceptor). Because resonance energy transfer is a distance-dependent phenomenon that increases in efficiency to the 6th power of the distance between the donor and the acceptor, assay systems are often designed to minimize donor-acceptor distances. However, the authors show that because of the R(6) relationship between transfer efficiency and sensitized emission lifetime, energy transfer can be difficult to measure in a time-gated manner when the donor-acceptor distance is small relative to the Förster radius. In such systems, the advantages inherent in time-resolved, ratiometric measurements are lost but can be regained by designing the system such that the average donor-acceptor distance is increased.
Vogel
Kurt W
KW
Invitrogen Drug Discovery Solutions, Madison, WI 53719, USA.
Vedvik
Kevin L
KL
D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; D010450:Endopeptidases / Q000032:analysis; D004735:Energy Transfer; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods* / Q000706:statistics & numerical data; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D028581:Lanthanoid Series Elements; D008962:Models, Theoretical
D005456:Fluorescent Dyes; D028581:Lanthanoid Series Elements; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human