Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate NPNT in samples. An antibody specific for NPNT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyNPNT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NPNT 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 NPNT bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Nephronectin contains 561 amino acids. In situ hybridization detected nephronectin in the developing mouse kidney and in several other tissues and organs. Expression appeared particularly prominent at epithelial-mesenchymal interfaces in tissues undergoing morphogenesis.
The deduced 565- and 582-amino acid proteins have calculated molecular masses of 61.9 and 64.0 kD, respectively. Both proteins contain an N-terminal signal sequence, followed by 5 EGF domains, and a C-terminal MAM domain. They also contain an RGD integrin-binding site, a potential glycosylation site, and a potential tyrosine phosphorylation site. EGF repeats 2, 4, and 5 contain Ca(2+)-binding EGF domains. In the longer variant, the 17 additional amino acids are inserted N-terminal to the first EGF domain.