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      Purified Mouse Anti-Human Granzyme A
      Purified Mouse Anti-Human Granzyme A
      Profile of permeabilized peripheral blood lymphocytes analyzed by flow cytometry (BDIS, San Jose, CA). Cells were collected, fixed, and permeabilized using the Cytofix/Cytoperm™ Kit (Cat. No. 554714) for 20 minutes at room temperature (RT), pelleted and washed twice with Perm/Wash Buffer™ (component of Cat. No. 554714). Cells were then stained with purified anti-human granzyme A antibody (Cat. No. 557449, 0.5 µg/test) or an IgG1 isotype control (Cat. No. 554121, clone MOPC-21) for 20-30 minutes at RT in the dark. Cells were then washed in Perm/Wash Buffer and stained with FITC labeled goat anti-mouse IgG/IgM, Cat. No. 555988, 0.25 µg/test) for 20-30 minutes at RT in the dark. Cells were washed once in Perm/Wash Buffer™, resuspended in wash buffer and analyzed by flow cytometry.
      Purified Mouse Anti-Human Granzyme A
      Profile of permeabilized peripheral blood lymphocytes analyzed by flow cytometry (BDIS, San Jose, CA). Cells were collected, fixed, and permeabilized using the Cytofix/Cytoperm™ Kit (Cat. No. 554714) for 20 minutes at room temperature (RT), pelleted and washed twice with Perm/Wash Buffer™ (component of Cat. No. 554714). Cells were then stained with purified anti-human granzyme A antibody (Cat. No. 557449, 0.5 µg/test) or an IgG1 isotype control (Cat. No. 554121, clone MOPC-21) for 20-30 minutes at RT in the dark. Cells were then washed in Perm/Wash Buffer and stained with FITC labeled goat anti-mouse IgG/IgM, Cat. No. 555988, 0.25 µg/test) for 20-30 minutes at RT in the dark. Cells were washed once in Perm/Wash Buffer™, resuspended in wash buffer and analyzed by flow cytometry.
      Product Details
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      BD Pharmingen™
      CTL tryptase; CTLA3; GZMA; GrA; HFSP; Hanukkah factor
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Human Cytotoxic T Cell Granule Proteins
      Flow cytometry (Routinely Tested), Immunoprecipitation (Reported)
      29 kDa
      0.5 mg/ml
      AB_396712
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      557449 Rev. 6
      Antibody Details
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      CB9

      The primary mechanism by which cytotoxic T cells eliminate virally infected cells is by granule exocytosis. The release of cytotoxic granule contents by cytotoxic T lymphocytes (CTL) triggers apoptotic target cell death. CTL granules contain a pore-forming protein, perforin, and a group of serine proteases called granzymes. In the classic model, perforins create holes in the target cell membrane, allowing entrance of the granzymes. Granzyme A and B are the predominant granzymes activated after CTL activation, but each act via an independent apoptotic pathway; granzyme B is activated immediately, while granzyme A acts hours later. Granzyme B has been shown to induce apoptosis and to cleave a number of substrates which are similar in specificity to those of the caspase family of proteinases. Granzyme B can cleave substrates, such as DNA-PKcs, and nuclear mitotic apparatus protein (NuMA). Furthermore, Granzyme B can also cleave substrates such as Bid and DFF45 in a caspase-independent fashion. Studies involving mice which are deficient in both granzyme A and B suggest a model whereby the granzyme B pathway may have evolved as the major apoptotic pathway with the granzyme A pathway acting as a backup. However, further research is needed to delineate the components of these distinct pathways. Clone CB9 recognizes human granzyme A. Purified human granzyme A was used as the immunogen.

      This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

      557449 Rev. 6
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      557449 Rev.6
      Citations & References
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      Development References (8)

      1. Andrade F, Roy S, Nicholson D, Thornberry N, Rosen A, Casciola-Rosen L. Granzyme B directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis. Immunity. 1998; 8(4):451-460. (Biology). View Reference
      2. Barry M, Heibein JA, Pinkoski MJ, et al. Granzyme B short-circuits the need for caspase 8 activity during granule-mediated cytotoxic T-lymphocyte killing by directly cleaving Bid. Mol Cell Biol. 2000; 20(11):3781-3794. (Biology). View Reference
      3. Beresford PJ, Kam CM, Powers JC, Lieberman J. Recombinant human granzyme A binds to two putative HLA-associated proteins and cleaves one of them. Proc Natl Acad Sci U S A. 1997; 94(17):9285-9290. (Immunogen: Immunoprecipitation). View Reference
      4. Beresford PJ, Xia Z, Greenberg AH, Lieberman J. Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation. Immunity. 1999; 10(5):585-594. (Clone-specific: Immunoprecipitation). View Reference
      5. Pinkoski MJ, Waterhouse NJ, Heibein JA, et al. Granzyme B-mediated apoptosis proceeds predominantly through a Bcl-2-inhibitable mitochondrial pathway. J Biol Chem. 2001; 276(15):12060-12067. (Biology). View Reference
      6. Sharif-Askari E, Alam A, Rheaume E, et al. Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation. EMBO J. 2001; 20(12):3101-3113. (Biology). View Reference
      7. Shresta S, Graubert TA, Thomas DA, Raptis SZ, Ley TJ. Granzyme A initiates an alternative pathway for granule-mediated apoptosis. Immunity. 1999; 10(5):595-605. (Biology). View Reference
      8. Trimble LA, Lieberman J. Circulating CD8 T lymphocytes in human immunodeficiency virus-infected individuals have impaired function and downmodulate CD3 zeta, the signaling chain of the T-cell receptor complex. Blood. 1998; 91(2):585-594. (Clone-specific: Flow cytometry). View Reference
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      557449 Rev. 6

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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