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Fluorescence resonance energy transfer (more commonly known as FRET) allows the detection of the approach between two molecules within several nanometers.
This distance-dependent interaction is sufficiently close to allow molecular reactions to occur and can provide information on potential interactions between labeled molecules within a cell.
Fluorescence is the light signal we can detect when a fluorophore absorbs energy at a specific wavelength and is excited to emit light at a higher wavelength as it returns to its ground state.
During FRET, a donor fluorophore becomes excited by a light source and transfers its energy to a nearby acceptor fluorophore. The acceptor fluorophore absorbs the energy to produce a detectable light emission signal. This process results in the loss of fluorescence of the donor and the gain of fluorescence of the acceptor, both of which can be measured.
For FRET to occur, several conditions must be met:
FRET is an extremely powerful method of identifying potential molecular interactions and can be used in techniques such as flow cytometry, immunocytochemistry, immunohistochemistry, and ELISA. FRET is also ideally suited to high-throughput screening (HTS) since it is simple, sensitive, and easily automated.
One popular use of FRET is to identify an interaction between two biomolecules, for example, the binding of a ligand to a receptor; a FRET signal can only be detected when the biomolecules are in close proximity by virtue of a binding event.
FRET relies on the use of high-quality labeled reagents. Depending on the intended assay setup, these could be antibodies, proteins, or peptides.
We have identified the following optimum FRET pairs that can be found across our antibody range, generated using Lightning-Link® conjugation kits or manufactured via our custom labeling services to offer full assay flexibility:
We offer a wide range of fluorescent-labeled antibodies, proteins and dyes, covering the spectrum from UV to far-infrared to help meet all your experimental needs.