What is difference between FRET and TR-FRET?

TR-FRET combines the low background aspect of TRF with the homogeneous assay format of FRET. The resulting assay provides an increase in flexibility, reliability, and sensitivity in addition to higher throughput and fewer false-positive/false-negative results. FRET involves two fluorophores, a donor and an acceptor.

Is Htrf the same as TR-FRET?

Homogeneous Time-Resolved Fluorescence (HTRF®) is a Time-Resolved FRET (TR-FRET) immunoassay developed by Cisbio. This technology combines Time-Resolved Fluorescence (TRF) detection with Fluorescence Resonance Energy Transfer (FRET) and is mainly used for drug screening in high-throughput.

Can you do FRET with antibodies?

​​FRET relies on the use of high quality labeled reagents. Depending on the intended assay setup these could be antibodies, proteins or peptides.

What is d2 Htrf?

Overview. Streptavidin has been labeled with d2. Biotin binds to Streptavidin with high affinity (Ka=1015M-1). The binding is rapid and stable, making it an ideal choice for use in a variety of assays such as enzyme assays, protein-protein binding assays and molecular biology assays.

How is lifetime fluorescence measured?

Fluorescence lifetime can be measured in either the frequency domain or the time domain. The time domain method involves the illumination of a sample (a cuvette, cells, or tissue) with a short pulse of light, followed by measuring the emission intensity against time.

What is TRF telomere?

Terminal Restriction Fragments (TRF) analysis is the oldest approach to analyze telomere lengths and remains the “gold standard” even in current studies. This technique relies on the fact that repeated minisatellite telomeric units do not contain target sites for restriction enzymes.

What parameters affect FRET?

The FRET efficiency depends on many physical parameters that can be grouped as: 1) the distance between the donor and the acceptor (typically in the range of 1–10 nm), 2) the spectral overlap of the donor emission spectrum and the acceptor absorption spectrum, and 3) the relative orientation of the donor emission …

What is Htrf used for?

HTRF (Homogeneous Time Resolved Fluorescence) is the most frequently used generic assay technology to measure analytes in a homogenous format, which is the ideal platform used for drug target studies in high-throughput screening (HTS).

What is the lifetime range of fluorescence?

Principles. The fluorescence lifetime is a measure of the time a fluorophore spends in the excited state before returning to the ground state by emitting a photon [1]. The lifetimes of fluorophores can range from picoseconds to hundreds of nanoseconds.

What does fluorescence lifetime depend on?

It is based on the fact that the fluorescence lifetime of a fluorophore depends on its molecular environment but not on its concentration. It can be applied in fluorescence microscopy where the local probe concentration cannot be controlled.