Hepatocyte growth factor (HGF),also known as Hepatopoietin A,is mitogenic for a variety of cell types,including endothelial and epithelial cells,melanocytes,and keratinocytes. It is identical to scatter factor,a fibroblast-derived soluble factor that promotes the dissociation of epithelial and vascular endothelial cell colonies in monolayer cultures by stimulating cell migration. HGF is secreted as an inactive single chain precursor and is converted to the heterodimeric active form by HGF activator.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human HGF BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human HGF concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:30pg/mlRange:125-8000pg/mlSpecificity:Recognizes human HGF.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h HGF monoclonal coating antibody is adsorbed onto microwells. HGF 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 HGF antibody is added and binds to HGF captured by the first antibody. Following incubation unbound Biotinylated anti-h HGF antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h HGF antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of HGF 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 HGF standard dilutions and HGF sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 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.