Granzyme B is a serine protease that in humans is encoded by the GZMB gene. Granzyme B is expressed by cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. CTL and NK cells share the remarkable ability to recognize specific infected target cells. They are thought to protect their host by inducing apoptosis of cells that bear on their surface nonself antigens, usually peptides or proteins resulting from infection by intracellular pathogens. The protein encoded by this gene is crucial for the rapid induction of target cell apoptosis by CTL in cell-mediated immune response. 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 Mouse Granzyme B. Sensitivity: <10pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Mouse Granzyme B. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse Granzyme B BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A monoclonal antibody from rat specific for Granzyme B has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: I21-S247) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Granzyme B 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 Mouse Granzyme B amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Mouse Gzmb antibody, 1x plate Lyophilized recombinant Mouse Gzmb standard, 2x10ng/tube Biotinylated anti-Mouse Gzmb 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.
