Hepatocyte growth factor activator, also known as HGFA, is a protein that in humans is encoded by the HGFAC gene. This gene encodes a member of the peptidase S1 protein family. The encoded protein is first synthesized as an inactive single-chain precursor before being activated to a heterodimeric form by endoproteolytic processing. It acts as serine protease that converts hepatocyte growth factor to the active form. Alternative splicing results in multiple transcript variants. Sample Type: Cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human HGFA. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human HGFA. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human HGFA BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for HGFA has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: Q36-S655) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for HGFA 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 HGFA amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human HGFAC antibody, 1x plate Lyophilized recombinant Human HGFAC standard, 2x10ng/tube Biotinylated anti-Human HGFAC antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer 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.
