TIMPs-1 through -4 regulate the activity of zinc metalloproteases known as MMPs，ADAMs and ADAMTSs. Structurally，TIMPs contain two domains. The N-terminal domain binds to the active site of mature metalloproteases via a 1：1 non-covalent interaction，blocking access of substrates to the catalytic site. In addition，The C-terminal domain of TIMP-1 and TIMP-2 binds to the hemopexin- like domain of pro-MMP-9 and pro-MMP-2，respectively. The latter binding is essential for the cell surface activation of MMP-2 by MMP-14.Sample Type：Cell culture supernatants，serum，plasma，and tissueIntended Use：The Human TIMP-1 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human TIMP-1 concentrations in cell culture supernatants，serum，plasma，and tissue.Sensitivity：30pg/mlRange：62.5-4000pg/mlSpecificity：Recognizes human TIMP-1.Sample Volume：100ul/wellTest Principle：This assay employs the Sandwich immunoassay technique. An anti-h TIMP-1 monoclonal coating antibody is adsorbed onto microwells. TIMP-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 TIMP-1 antibody is added and binds to TIMP-1 captured by the first antibody. Following incubation unbound Biotinylated anti-h TIMP-1 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h TIMP-1 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of TIMP-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 TIMP-1 standard dilutions and TIMP-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.
