CD40:CD40L TR-FRET Assay Kit

TR-FRET

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  The CD40:CD40L TR-FRET Assay Kit is designed to measure binding activity of CD40 (cluster of differentiation 40) to CD40L (CD40 ligand) for screening and profiling applications using TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer). It utilizes Terbium-labeled donor and a labeled acceptor to complete the TR-FRET pairing. The CD40:CD40L TR-FRET Assay Kit comes in a convenient 384-well format, with enough recombinant biotinylated CD40 (amino acids 21-193), recombinant CD40L (amino acids 113-261), Tb-Labeled Donor and Dye-Labeled Acceptor and assay buffer for 384 reactions.

  CD40:CD40L TR-FRET Assay Kit schematic.
  A sample containing terbium-labeled donor, dye-labeled acceptor, CD40, CD40L, and an inhibitor is incubated for 90 minutes. The fluorescence intensity is then measured using a fluorescence reader. In the presence of binding of CD40 to CD40L, energy transfer occurs due to the proximity of the donor and acceptor. Disruption of the binding results in decrease of energy transfer. Fluorescence intensity at λ=665 nm corresponds directly to the binding of CD40 to CD40L.