Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate VWA3A in samples. An antibody specific for VWA3A has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyVWA3A present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for VWA3A is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of VWA3A bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:vWA3A,Contains 2 VWFA domains, Human WARP shares 79% amino acid identity with mouse Warp. RT-PCR of mRNA from various mouse tissues and cell lines detected Warp expression only in chondrocytes. Northern blot analysis confirmed expression of Warp in chondrocytes isolated from newborn mouse rib cartilage.
Western blot analysis of transfected human embryonic kidney cells detected a 48-kD protein in both cell layer and media fractions. The majority of the protein was detected in the medium, indicating that WARP is efficiently secreted. Under nonreducing conditions, a 102-kD form was detected, suggesting that WARP forms a disulfide-bonded homodimer. N-glycosidase digestion led to a mobility shift from 48 to 45 kD, indicating that WARP has 1 or more N-linked oligosaccharide chains.
