TR-FRET

Left Side (FRET Occurs): In the presence of an active enzyme-ligand complex, the fluorescence donor [F(d)] and acceptor [F(a)] are in close proximity, which allows energy transfer (FRET) from the donor to the acceptor. This transfer generates a detectable signal, indicating a successful interaction between the enzyme and ligand.

Inhibitor Added: When an inhibitor molecule (I) is introduced, it binds to the enzyme, disrupting the interaction between the donor and acceptor. This results in reduced or no FRET signal, as the inhibitor prevents the enzyme-ligand binding.

Right Side (No FRET): Without interaction between the enzyme and ligand, the donor and acceptor are too far apart for energy transfer, so no FRET signal is produced.

Its high sensitivity also enables experiments with minimal sample volumes, which is especially valuable when working with rare or costly biological materials. TR-FRET’s versatility spans a wide range of applications, from studying receptor-ligand interactions to examining enzyme-substrate dynamics and protein conformational changes, offering researchers a powerful tool for understanding complex molecular interactions. 

Another significant advantage of TR-FRET is its resistance to photobleaching, thanks to its lanthanide-based donors. This feature ensures stable fluorescence signals over extended periods, allowing for more consistent data collection, even in assays requiring prolonged measurements or repeated readings. The technique’s reliability and sensitivity, combined with its low background interference and adaptability to various experimental needs, make it essential for fields such as drug discovery, cellular biology, and biochemical research.