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      PE Rat Anti-Mouse Granzyme C
      PE Rat Anti-Mouse Granzyme C
      Flow cytometric analysis of Granzyme C expression in activated Mouse splenic leucocytes. C57BL/6 mouse splenic leucocytes were activated by culture with Recombinant Mouse IL-15 protein (100 ng/ml) for 4 days (37°C). The cells were harvested, washed, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] then stained with BD Horizon™ BUV395 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 564144) and BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406). The cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse Granzyme C antibody (Cat. No. 568735; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of Granzyme C (or Ig Isotype control staining) versus NK-1.1 was derived from gated events with the forward and side light- scatter characteristics of viable (Fixable Viability Stain 510-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific
      PE Rat Anti-Mouse Granzyme C
      Flow cytometric analysis of Granzyme C expression in activated Mouse splenic leucocytes. C57BL/6 mouse splenic leucocytes were activated by culture with Recombinant Mouse IL-15 protein (100 ng/ml) for 4 days (37°C). The cells were harvested, washed, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] then stained with BD Horizon™ BUV395 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 564144) and BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406). The cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse Granzyme C antibody (Cat. No. 568735; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of Granzyme C (or Ig Isotype control staining) versus NK-1.1 was derived from gated events with the forward and side light- scatter characteristics of viable (Fixable Viability Stain 510-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific
      Product Details
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      BD Pharmingen™
      Gzmc; cytotoxic cell protease 2; CCP2; Ctla5; Ctla-5
      Mouse (QC Testing)
      Rat IgG1, κ
      Purified Recombinant Mouse Granzyme C Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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 the dye under optimum conditions and unconjugated antibody and free dye were removed.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      6. An isotype control should be used at the same concentration as the antibody of interest.
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      568735 Rev. 1
      Antibody Details
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      SFC1D8.rMAb

      The SFC1D8.rMAb is a recombinant Rat IgG1 κ monoclonal antibody with VH and VL regions derived from SFC1D8 hybridoma cells that secrete Armenian Hamster IgG antibodies specific for Mouse Granzyme C. Granzyme C is also known as Cytotoxic cell protease 2 (CCP2) or Ctla5 (Ctla-5). Granzyme C is a serine protease that is encoded by Gzmc (granzyme C) which belongs to the peptidase S1 family. Granzyme C is found in the cytotoxic granules of cytotoxic T lymphocyte and natural killer (NK) effector cells. Upon recognition of target cells, these effector cells can exocytose Granzyme C which induces apoptosis of target cells leading to their externalization of phosphatidylserine, nuclear condensation, mitochondrial swelling, and the single-stranded DNA nicks. Granzyme C can reportedly support cytotoxic T lymphocyte-mediated killing of target cells in the absence of Granzyme A or B.

      568735 Rev. 1
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      568735 Rev.1
      Citations & References
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      View product citations for antibody "568735" on CiteAb

      Development References (9)

      1. Cai SF, Fehniger TA, Cao X, et al. Differential expression of granzyme B and C in murine cytotoxic lymphocytes.. J Immunol. 2009; 182(10):6287-97. (Immunogen: ELISA, Flow cytometry). View Reference
      2. Cao X, Cai SF, Fehniger TA, et al. Granzyme B and perforin are important for regulatory T cell-mediated suppression of tumor clearance.. Immunity. 2007; 27(4):635-46. (Biology). View Reference
      3. Garcia-Sanz JA, MacDonald HR, Jenne DE, Tschopp J, Nabholz M. Cell specificity of granzyme gene expression.. J Immunol. 1990; 145(9):3111-8. (Biology). View Reference
      4. Getachew Y, Stout-Delgado H, Miller BC, Thiele DL. Granzyme C supports efficient CTL-mediated killing late in primary alloimmune responses.. J Immunol. 2008; 181(11):7810-7. (Biology). View Reference
      5. Janas ML, Groves P, Kienzle N, Kelso A. IL-2 regulates perforin and granzyme gene expression in CD8+ T cells independently of its effects on survival and proliferation.. J Immunol. 2005; 175(12):8003-10. (Biology). View Reference
      6. Jenne D, Rey C, Masson D, et al. cDNA cloning of granzyme C, a granule-associated serine protease of cytolytic T lymphocytes.. J Immunol. 1988; 140(1):318-23. (Biology). View Reference
      7. Johnson H, Scorrano L, Korsmeyer SJ, Ley TJ. Cell death induced by granzyme C.. Blood. 2003; 101(8):3093-101. (Biology). View Reference
      8. Kelso A, Costelloe EO, Johnson BJ, Groves P, Buttigieg K, Fitzpatrick DR. The genes for perforin, granzymes A-C and IFN-gamma are differentially expressed in single CD8(+) T cells during primary activation.. Int Immunol. 2002; 14(6):605-13. (Biology). View Reference
      9. Revell PA, Grossman WJ, Thomas DA, et al. Granzyme B and the downstream granzymes C and/or F are important for cytotoxic lymphocyte functions.. J Immunol. 2005; 174(4):2124-31. (Biology). View Reference
      View All (9) View Less
      568735 Rev. 1

      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.