Description
Perforin has a key role in cell-mediated cytotoxicity. It is a 70 kDa cytolytic protein that is expressed in the cytoplasmic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. CTLs are involved in eliminating virally infected cells, in anti-tumor immune responses, in allograft rejections, and in some autoimmune diseases. NK cells are important for tumor surveillance and destruction and are involved in allograft rejections. Cytotoxic cells release the contents of their cytotoxic granules, including perforin upon recognition of their target cell. In the presence of calcium, perforin forms transmembrane channels or pores in the membrane of the target cell leading to a cell death that resembles apoptosis. The ability to detect perforin-positive cells with specific antibody should be useful in identifying and understanding perforin-mediated reactions. Clone δG9 reacts with human and bovine perforin. It does not cross-react with mouse perforin. Purified granules from the human lymphoma cell line YT were used as immunogen. Clone δG9 was initially characterized by immunoprecipitation and immunohistochemistry of frozen tissue sections. The antibody stains scattered lymphocytes in red pulp of spleen, and scattered infiltrated lymphocytes in lymphoma.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
6. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
