Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PLBD1 in samples. An antibody specific for PLBD1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPLBD1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PLBD1 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 PLBD1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:PLBD1, Belongs to the phospholipase B-like family. A phospholipase B (PLB) precursor was purified from normal human granulocytes using Sephadex G-75, Mono-S cation-exchange and hydroxyapatite columns. The native protein needed modifications to acquire deacylation activity against phospholipids including phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine and lysophospholipids. The enzyme revealed a PLB nature by removing fatty acids from both the sn-1 and sn-2 positions of phospholipids. The enzyme is active at a broad pH range with an optimum of 7.4. Immunoblotting of neutrophil postnuclear supernatant using antibodies against the 42 kDa fragment detected a band at a molecular mass of 42 kDa, indicating a neutrophil origin of the novel PLB precursor.
