Principle of the Assay: This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to GzmB. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for GzmB and Avidin-Horseradish Peroxidase (HRP) conjugate is added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain GzmB, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The OD value is proportional to the concentration of GzmB. You can calculate the concentration of GzmB in the samples by comparing the OD of the samples to the standard curve.
NCBI and Uniprot Product Information
NCBI Description
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 encoded by this gene is crucial for the rapid induction of target cell apoptosis by CTL in cell-mediated immune response. [provided by RefSeq, Jul 2008]
Uniprot Description
GZMB: This enzyme is necessary for target cell lysis in cell- mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis. By staphylococcal enterotoxin A (SEA) in peripheral blood leukocytes. Inactivated by the serine protease inhibitor diisopropylfluorophosphate. Belongs to the peptidase S1 family. Granzyme subfamily.
Protein type: Protease; EC 3.4.21.79; Apoptosis
Chromosomal Location of Human Ortholog: 14q11.2
Cellular Component: intracellular membrane-bound organelle; membrane; cytoplasm; immunological synapse; cytosol; nucleus
Molecular Function: protein binding; serine-type peptidase activity; serine-type endopeptidase activity
Biological Process: Notch signaling pathway; induction of apoptosis by granzyme; apoptosis; natural killer cell mediated cytotoxicity; cytolysis; protein processing; immune response; proteolysis