Description: One of the major mechanisms of cell-mediated cytotoxicity involves exocytosis of cytoplasmic granules from the effector toward the target cell. The granules contain a number of proteins, including the pore-forming protein perforin and a family of serine proteases called granzymes, including Granzyme B. Granzyme B is present mainly in the granules of CD8+ CTL and natural killer (NK) cells and mediates the lethal hit that kills virus infected and tumorigenic cells. Measurement of release of Granzyme B in response to the appropriate target is useful for evaluating cell-mediated cytotoxicity. The Granzyme B ELISPOT assay is a valuable non-radioactive alternative to the 51Cr-release killing cell assays for measuring antigen-specific CTL cytotoxicity.
This Granzyme B ELISPOT Ready-SET-Go! reagent set contains the necessary reagents for performing enzyme linked immunosorbent spot (ELISPOT) assays for high resolution frequency analysis of Granzyme B-secreting cells. This ELISPOT reagent set is pre-titrated for optimal spot development.
Components
Capture Antibody. Pre-titrated, Functional Grade (low endotoxin) purified antibody Detection Antibody. Pre-titrated, biotin-conjugated antibody ELISA/ELISPOT Coating Buffer. This Ready-Set-Go! ELISPOT Set may contain ELISA/ELISPOT Coating Buffer Powder (Reconstitute to 1L with dH20 and filter (0.22 uM)) or 10X PBS ELISPOT Coating Buffer (Dilute 1 part 10X Buffer into 9 parts dH20 and filter with 0.22 uM). Assay Diluent. 5X concentrated Detection enzyme. Pre-titrated Avidin-HRP Certificate of Analysis. Lot-specific instructions for dilution of antibodies and enzyme
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