Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate sPLA2 in samples. An antibody specific for sPLA2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anysPLA2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for sPLA2 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 sPLA2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Phospholipases A2 (PLA2s) hydrolyze the sn-2 fatty acid acyl ester bond of phosphoglycerides, releasing free fatty acids and lysophospholipids. PLA2G2A belongs to PLA2 group II, which consists of extracellular enzymes with low molecular masses that require calcium ions for catalysis . The deduced protein contains a 20-amino acid signal sequence, followed by a 124-amino acid mature peptide that has a calculated molecular mass of about 14 kD. The protein has characteristics of the type II class of PLA2s, including conserved catalytic residues and a Ca(2+)-binding loop. Northern blot analysis detected a 0.8-kb transcript in inflamed human synovial tissue and in peritoneal exudate cells from a peritonitis patient, but not in 2 activated human promyelocytic cell lines.
