Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate NID2 in samples. An antibody specific for NID2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyNID2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NID2 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 NID2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:NID2 protein, 46% identical to NID1, contains a 30-residue signal peptide, 49 primarily central cys residues, 5 potential N-linked glycosylation sites, 2 tyr residues in a potential O-sulfation sequence, and a YGD rather than an RGD cell adhesion sequence.SDS-PAGE analysis showed that NID2 is expressed as a 200-kD protein, larger than the calculated mass of 148 kD, presumably due to oligosaccharide substitution as indicated by hexosamine analysis. Northern blot analysis revealed ubiquitous expression of a 5.5-kb NID2 transcript that was strongest in heart and placenta, moderate in pancreas, kidney, and skeletal muscle, and weakest in brain. Immunoblot analysis detected expression of NID2 in muscle, heart, placenta, kidney, skin, and testis, with weaker expression in liver and brain.
