BD Horizon™ R718 Rat Anti-Mouse Granzyme C
Clone SFC1D8.rMAb (RUO)
Multicolor flow cytometric analysis of Granzyme C expression in activated Mouse splenic leukocytes. C57BL/6 Mouse splenocytes 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] and then stained with APC Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 550627) and BD Horizon™ Fixable Viability Stain 780 (Cat. No. 565388). The cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed, permeabilized with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ R718 Rat IgG1, κ Isotype Control (Cat. No. 566948; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Granzyme C antibody (Cat. No. 569825/569900; Right Plot) at 1 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 780-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
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Multicolor flow cytometric analysis of Granzyme C expression in activated Mouse splenic leukocytes. C57BL/6 Mouse splenocytes 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] and then stained with APC Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 550627) and BD Horizon™ Fixable Viability Stain 780 (Cat. No. 565388). The cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed, permeabilized with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ R718 Rat IgG1, κ Isotype Control (Cat. No. 566948; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Granzyme C antibody (Cat. No. 569825/569900; Right Plot) at 1 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 780-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
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
14940
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. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
R718 Rat IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
566948
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Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
569825 Rev. 1
Antibody Details
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.
569825 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
569825 Rev.1
Citations & References
View product citations for antibody "569825" on CiteAb
Development References (9)
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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, Intracellular Staining/Flow Cytometry). View Reference
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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
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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
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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
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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
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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
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Johnson H, Scorrano L, Korsmeyer SJ, Ley TJ. Cell death induced by granzyme C.. Blood. 2003; 101(8):3093-101. (Biology). View Reference
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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
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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
569825 Rev. 1
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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.