Description
Principle of the Assay::The Rat CD265 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Rat CD265 in Cell Culture Supernatants, Serum. This assay employs an antibody specific for Rat CD265 coated on a 96-well plate. Standards and samples are pipetted into the wells and CD265 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat CD265 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of CD265 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background/Introduction: Receptor Activator of Nuclear Factor kappa B (RANK), also known as TRANCE Receptor, is a type I membrane protein that is expressed on the surface of osteoclasts and is involved in their activation upon ligand binding. RANK is a recently described TNF receptor family member, and its ligand, RANKL, promote survival of dendritic cells and differentiation of osteoclasts. RANK contains 383 amino acids in its intracellular domain (residues 234-616), which contain three putative TRAF-binding domains (termed I, II, and III). RANK interacts with various TRAFs through distinct motifs and activates NF-kappaB via a novel TRAF6 interaction motif, which then activates NIK, thus leading to NF-kappaB activation, whereas RANK most likely activates JNK through a TRAF2-interacting region in RANK. The standard in this kit is recombinant human RANK with the sequence of Q29-G213 aa. It is a dipolymer which compose of two chains, and the molecular weight of each is 48kda