Hepatocyte growth factor (HGF) is the most potent mitogen for mature parenchymal hepatocytes in primary culture, and seems to be a hepatotrophic factor that acts as a trigger for liver regeneration after partial hepatectomy and liver injury. HGF has a relative molecular mass (Mr) of 82,000 and is a heterodimer composed of a large alpha-subunit of Mr 69,000 and a small beta-subunit of Mr 34,000.1 The protein consists of 728aa residues, including a possible signal peptide at the N-terminus.2 HGF may serve as a paracrine mediator to control placental development and growth.3 This growth factor may play an important role as a paracrine mediator of the proliferation of melanocytes and endothelial cells, as well as cells of epithelial origin.4 The gene encoding the human HGF is assigned to human chromosome 7. Sample Type: Cell culture supernates, serum and plasma (EDTA, citrate) Intended Use: For quantitative detection of rat HGF in cell culture supernates, serum and plasma(EDTA, citrate). Sensitivity: <10pg/ml Range: 156-10,000pg/ml Specificity: Natural and recombinant rat HGF Test Principle: This rat HGF BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for HGF has been precoated onto 96-well plates. Standards(NSO, Q33-R495(alpha) & V496-L728(beta)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for HGF 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 rat HGF amount of sample captured in plate. Kit Components: 96-well plate precoated with rat HGF antibody, 1 Lyophilized recombinant rat HGF standard, 10ng/tube*2 Biotinylated rat HGF antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml 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.
