Granzyme A is a protein that in humans is encoded by the GZMA gene. Cytolytic T lymphocytes (CTL) and natural killer (NK) cells share the remarkable ability to recognize, bind, and lyse specific target cells. They are thought to protect their host by lysing cells bearing on their surface nonself antigens, usually peptides or proteins resulting from infection by intracellular pathogens. The protein described here is a T cell- and natural killer cell-specific serine protease that may function as a common component necessary for lysis of target cells by cytotoxic T lymphocytes and natural killer cells. GZMA induces caspase-independent apoptosis in a characteristic manner, except it causes a distinctive form of DNA damage: single-stranded DNA nicking. A target of GZMA is the SET complex, including HMGB2 and ANP32A. Sample Type: Cell culture supernates, serum and plasma (heparin, EDTA) Intended Use: For quantitative detection of human Granzyme A in cell culture supernates, serum and plasma (heparin, EDTA). Sensitivity: <10pg/ml Range: 15.6-1000pg/ml Specificity: Natural and recombinant human Granzyme A Test Principle: This human Granzyme A BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Granzyme A has been precoated onto 96-well plates. Standards(NSO) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Granzyme A 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 Granzyme A amount of sample captured in plate. Kit Components: 96-well plate precoated with human Granzyme A antibody, 1 Lyophilized recombinant human Granzyme A standard, 10ng/tube*2 Biotinylated human Granzyme A 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.
