TFPI(Tissue factor pathway inhibitor), also known as EPI, LACI or TFPI1, is a single-chain polypeptide which can reversibly inhibit Factor Xa (Xa) and Thrombin (Factor IIa). The gene for TFPI is located on chromosome 2q31-q32.1, and has nine exons which span 70 kb. TFPI consists of a highly negatively-charged amino-terminus, three tandemly-linked Kunitz domains, and a highly positively-charged carboxy-terminus. TFPI contributes significantly to the inhibition of Xa in vivo, despite being present at concentrations of only 2.5 nM. Sample Type: Cell culture supernates, serum and plasma (heparin, EDTA) Intended Use: For quantitative detection of mouse TFPI in cell culture supernates, serum and plasma (heparin, EDTA). Sensitivity: <10pg/ml Range: 156-10,000pg/ml Specificity: Natural and recombinant mouse TFPI Test Principle: This mouse TFPI BioAssay(TM) ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for TFPI has been precoated onto 96-well plates. Standards(NSO, L29-N306) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TFPI is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. HRP substrate TMB are used to visualize HRP enzymatic reaction. TMB is 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 mouse TFPI amount of sample captured in plate. Kit Components: 96-well plate precoated with mouse TFPI antibody, 1 Lyophilized recombinant mouse TFPI standard, 10ng/tube*2 Biotinylated mouse TFPI 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.
