Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PLA2G12B in samples. An antibody specific for PLA2G12B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPLA2G12B present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PLA2G12B 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 PLA2G12B bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:The predicted 195-amino acid human protein shares 90% identity with the mouse protein and 40% identity with human PLA2G12A. It contains a 19-amino acid signal peptide and 14 cysteines, but it lacks N-glycosylation sites and has a leucine in place of the canonical histidine in its active site.
PLA2G12B is conserved in vertebrates, but not in invertebrates. SDS-PAGE revealed a 20-kD PLA2G12B protein, consistent with the calculated molecular mass of the mature protein. Northern blot analysis detected 1.4- and 6.5-kb PLA2G12B transcripts in liver, kidney, skeletal muscle, and heart. Expression of PLA2G12B was significantly weaker in tumors of kidney, liver, and small intestine compared with matched normal tissues. PLA2G12B expression varied in cancer cell lines.
