The human serpin superfamily consists of at least 35 members that target not only serine proteases,but also selected cysteine proteases and non-protease proteins. Serpins bind the protease active site resulting in a major conformational rearrangement that traps the enzyme in a covalent acyl-enzyme intermediate. As protease inhibitors,serpins have an array of functions including regulating blood clotting,the complement pathway,extracellular matrix remodeling,and cell motility. They are also involved in activities that extend beyond their ability to inhibit proteases. For instance,they may also regulate blood pressure,angiogenesis,or act as storage/transport proteins.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human Serpin E1/PAI-1 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human Serpin E1/PAI-1 concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:70pg/mlRange:156-10000pg/mlSpecificity:Recognizes human Serpin E1/PAI-1.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h Serpin E1/PAI-1 monoclonal coating antibody is adsorbed onto microwells. Serpin E1/PAI-1 present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h Serpin E1/PAI-1 antibody is added and binds to Serpin E1/PAI-1 captured by the first antibody. Following incubation unbound Biotinylated anti-h Serpin E1/PAI-1 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h Serpin E1/PAI-1 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of Serpin E1/PAI-1 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven Serpin E1/PAI-1 standard dilutions and Serpin E1/PAI-1 sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x6Standard: 1xVialBiotinylated antibody: 1xVialStreptavidin-HRP: 1xVialAssay Buffer (25ml/Vial): 1xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x6mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 2xstripsInstruction: 1xStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product,centrifuge the original vial after thawing and prior to removing the cap.