Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PNLIPRP2 in samples. An antibody specific for PNLIPRP2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPNLIPRP2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PNLIPRP2 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 PNLIPRP2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:PNLIPRP2 has 65% amino acid sequence identity with PNLIP, 63% identity with PNLIPRP1, and 20 to 30% identity with human gastric lipase, lipoprotein lipase, and hepatic lipase. Western blot analysis of recombinant PNLIPRP2 expressed in mammalian cells detected a secreted, approximately 50-kD protein; the authors concluded that PNLIPRP2 was glycosylated. In a standard pancreatic lipase assay, recombinant PNLIPRP2 showed lipolytic activity that was marginally increased by the addition of colipase and was inhibited by the lipase inhibitor orlistat. Northern blot analysis of pancreas RNA detected major 2.3- and 1.8-kb PNLIPRP2 transcripts. The expression level of PNLIPRP2 in pancreas was approximately 24-fold lower and 6-fold lower than the expression levels of PNLIP and PNLIPRP1, respectively.
