Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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 Powder. Reconstitute to 1L with dH20 and filter (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
Applications Reported
For research use only, not for diagnostic or therapeutic use.
References
Shafer-Weaver, K., et al. 2003. The Granzyme B ELISPOT assay: an alternative to the 51Cr-release assay for monitoring cell-mediated cytotoxicity. J. Translational Med. 1: 14. Rininsland, F., et al. 2000. Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity. J Immunol Meth. 240:143-155.
Protocol
To view protocol, please download the TDS by clicking the PDF button above.