Trypsin is a serine protease from the PA clan superfamily, found in the digestive system of many vertebrates, where it hydrolyses proteins. In the duodenum, trypsin catalyzes the hydrolysis of peptide bonds, breaking down proteins into smaller peptides. The peptide products are then further hydrolyzed into aa via other proteases, rendering them available for absorption into the blood stream. Tryptic digestion is a necessary step in protein absorption as proteins are generally too large to be absorbed through the lining of the small intestine. Additionally, Trypsin is produced as the inactive zymogen trypsinogen in the pancreas. When the pancreas is stimulated by cholecystokinin, it is then secreted into the first part of the small intestine (the duodenum) via the pancreatic duct. Once in the small intestine, the enzyme enteropeptidase activates trypsinogen into trypsin by proteolytic cleavage. Auto catalysis can happen with trypsin using trypsinogen as the substrate. This activation mechanism is common for most serine proteases, and serves to prevent autodegradation of the pancreas. Sample Type: Cell culture supernates, serum, plasma (Heparin). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human Pan Trypsin. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human Pan Trypsin. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human Pan Trypsin BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for Pan Trypsin has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: A16-S247) and test samples are added to the wells, a biotinylated detection specific for Pan Trypsin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human Pan Trypsin amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human PRSS3 antibody, 1x plate Lyophilized recombinant Human PRSS3 standard, 2x10ng/tube Biotinylated anti-Human PRSS3 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
