Description
Principle of the assay: human Prostasin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Prostasin has been precoated onto 96-well plates. Standards(CHO, A33-G319) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Prostasin 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 Prostasin amount of sample captured in plate.
Background: Prostasin is a protein that in humans is encoded by the PRSS8 gene. It is a single-copy gene and mapped to 16p11.2. Prostasin is likely to be the more physiologically relevant protease. This gene encodes a trypsinogen, which is a member of the trypsin family of serine proteases. Coexpression of Prostasin with either Xenopus or rat ENaC in Xenopus oocytes resulted in a 60% to 80% increase in amiloride-sensitive sodium currents, and that the addition of aprotinin, a serine protease inhibitor, completely prevented this activation. This enzyme is highly expressed in prostate epithelia and is one of several proteolytic enzymes found in seminal fluid. The proprotein is cleaved to produce a light chain and a heavy chain which are associated by a disulfide bond. It is active on peptide linkages involving the carboxyl group of lysine or arginine