Principle of HTRF®
HTRF® is based on Time-Resolved Fluorescence (TRF) and Fluorescence Resonance Energy Transfer (FRET).
In HTRF® the lanthanide chelate is used as donor and is either Europium cryptate (Eu3+ cryptate) or Lumi4™-Tb (Tb2+ cryptate), and the acceptor is either XL665, a modified allophycocyanin, or d2, a second generation of acceptor. In addition, HTRF® implements a ratiometric measurement that uses both the emission wavelength of the donor and of the acceptor to correct for well-to-well variability and signal quenching from assay components and media, that is absent in other TR-FRET assays.
Applications of HTRF®
HTRF® is used to measure molecular interactions in many research areas, including:
- Protein-Protein Interactions
- GPCRs
- Biomarkers
- Phosphoproteins
- Kinases
- Cytokines
- Epigenetics
Application Notes related to HTRF®
cAMP/IP-One HTplex HTRF® assay with Mithras cAMP/IP-One HTplex cell-based experiment performed on Mithras LB 940 Multimode Microplate Reader using HTRF® technology
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Microplate Readers recommended for HTRF®
A microplate reader with the following characteristics is recommended:
- Fluorescence measurement using filters
- Flash lamp as light source
- Detector with extended spectral range (above 650 nm)
All microplate readers below are recommended for HTRF®.
HTRF® is a registered trademark of Cisbio Bioassays.