Carboxypeptidase B2 (CPB2), also known as carboxypeptidase U (CPU), plasma carboxypeptidase B (pCPB) or thrombin-activatable fibrinolysis inhibitor (TAFI), is an enzyme that, in humans, is encoded by the gene CPB2. CPB2 is synthesized by the liver and circulates in the plasma as a plasminogen- bound zymogen. When it is activated by proteolysis at residue Arg92 by the thrombin/thrombomodulin complex, CPB2 exhibits carboxypeptidase activity. Activated CPB2 reduces fibrinolysis by removing the fibrin C-terminal residues that are important for the binding and activation of plasminogen. 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 TAFI/CPB2. Sensitivity: <15pg/ml Range: 156pg/ml-10,000pg/ml Specificity: Recognizes natural and recombinant Human TAFI/CPB2. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human TAFI/CPB2 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for TAFI/CPB2 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: M1-V423) and test samples are added to the wells, a biotinylated detection specific for TAFI/CPB2 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 TAFI/CPB2 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human CPB2 antibody, 1x plate Lyophilized recombinant Human CPB2 standard, 2x10ng/tube Biotinylated anti-Human CPB2 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.
