What is TR-fret used for?
TRF and TR-FRET (HTRF) assays enable the analysis of molecular interactions in biochemical processes and are widely used to study kinase assays, cellular signaling pathways, protein-protein interactions, DNA-protein interactions, cell cytotoxicity and receptor-ligand binding.
Does FRET use antibodies?
A typical TR-FRET immunoassay uses two antibodies for sandwich detection. Each recognizes a distinct epitope on the target antigen, yet while one antibody (the donor) is conjugated to a lanthanide, the other (the acceptor) is conjugated to a small molecule fluorescent dye.
What is HTRF assay?
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 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.
What is time-resolved fluorescence?
Time-resolved fluorescence spectroscopy is a spectroscopy technique used to monitor interactions between molecules and motions that occur in the short periods. The ability to measure changes in the picosecond or nanosecond time range makes it a useful technique in biomolecular structure analysis and dynamics.
What is difference between FRET and TR fret?
What is FRET efficiency?
FRET efficiency, defined as the proportion of the donor molecules that have transferred excitation state energy to the acceptor molecules, increases with decreasing intermolecluar distance (typically over the range 1–10 nm for fluorescent proteins).
What is Bret assay?
The bioluminescence resonance energy transfer (BRET) assay can be used as an indicator of molecular approximation and/or interaction. A significant resonance energy transfer signal is generated when the acceptor, having the appropriate spectral overlap with the donor emission, is approximated with the donor.
What are FRET sensors?
FRET sensors report changes in energy transfer between a donor and an acceptor fluorescent protein that occur when an attached sensor domain undergoes a change in conformation in response to ligand binding.
What determines FRET efficiency?
The FRET efficiency can be calculated by subtracting the donor intensity in the presence of the acceptor from its intensity after photobleaching the acceptor, and then normalizing this value to the donor intensity after bleaching.
What is delfia assay?
Assay principle DELFIA® (dissociation-enhanced lanthanide fluorescence immunoassay) is a time-resolved fluorescence (TRF) intensity technology. Assays are designed to detect the presence of a compound or biomolecule using lanthanide chelate labeled reagents, separating unbound reagent using wash steps.