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Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
Companion Products
The OX-8 monoclonal antibody specifically binds to the hinge-like membrane-proximal domain of the 32 kDa α chain of the CD8 differentiation antigen. A truncated CD8 α' isoform has not been detected in the rat. The CD8 α and β chains (CD8a and CD8b, respectively) form a heterodimer on the surface of most thymocytes and a subpopulation of mature T lymphocytes (i.e., MHC class I-restricted T cells, including most T suppressor/cytotoxic cells). Intestinal intrapithelial lymphocytes, many CD8+ T cells of athymic rats, many activated CD4+ T cells, and most NK cells express CD8a without CD8b. It has been suggested that the expression of the CD8a/CD8b heterodimer is restricted to thymus-derived T lymphocytes. OX-8 antibody does not react with resting CD4+ T helper cells. CD8 is an antigen coreceptor on the T-cell surface which interacts with MHC class I molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase Ick. Macrophages have also been reported to express CD8 α and β chains, which are involved in signal transduction. Soluble OX-8 mAb partially blocks in vitro MLR and CTL activity.
The antibody was conjugated to BD Horizon™ BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (15)
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Barclay AN. The localization of populations of lymphocytes defined by monoclonal antibodies in rat lymphoid tissues. J Immunol. 1981; 42(4):593-600. (Clone-specific: Immunohistochemistry). View Reference
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Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
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Brideau RJ, Carter PB, McMaster WR, Mason DW, Williams AF. Two subsets of rat T lymphocytes defined with monoclonal antibodies.. Eur J Immunol. 1980; 10:609-615. (Immunogen: Flow cytometry). View Reference
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Classon BJ, Brown MH, Garnett D, et al. The hinge region of the CD8 alpha chain: structure, antigenicity, and utility in expression of immunoglobulin superfamily domains. Int Immunol. 1992; 4(2):215-225. (Clone-specific). View Reference
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Hirji N, Lin TJ, Befus AD. A novel CD8 molecule expressed by alveolar and peritoneal macrophages stimulates nitric oxide production. J Immunol. 1997; 158(4):1833-1840. (Clone-specific: Stimulation). View Reference
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Hirji N, Lin TJ, Bissonnette E, Belosevic M, Befus AD. Mechanisms of macrophage stimulation through CD8: macrophage CD8alpha and CD8beta induce nitric oxide production and associated killing of the parasite Leishmania major. J Immunol. 1998; 160(12):6004-6011. (Clone-specific: Stimulation). View Reference
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Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
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Johnson P, Gagnon J, Barclay AN, Williams AF. Purification, chain separation and sequence of the MRC OX-8 antigen, a marker of rat cytotoxic T lymphocytes. EMBO J. 1985; 4(10):2539-2545. (Clone-specific: Immunoaffinity chromatography). View Reference
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Kuhnlein P, Park JH, Herrmann T, Elbe A, Hunig T. Identification and characterization of rat gamma/delta T lymphocytes in peripheral lymphoid organs, small intestine, and skin with a monoclonal antibody to a constant determinant of the gamma/delta T cell receptor. J Immunol. 1994; 153(3):979-986. (Biology). View Reference
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Mason DW, Arthur RP, Dallman MJ, Green JR, Spickett GP, Thomas ML. Functions of rat T-lymphocyte subsets isolated by means of monoclonal antibodies. Immunol Rev. 1983; 74:57-82. (Clone-specific: Blocking). View Reference
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Mitnacht R, Bischof A, Torres-Nagel N, Hunig T. Opposite CD4/CD8 lineage decisions of CD4+8+ mouse and rat thymocytes to equivalent triggering signals: correlation with thymic expression of a truncated CD8 alpha chain in mice but not rats. J Immunol. 1998; 160(2):700-707. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Scriba A, Grau V, Steiniger B. Phenotype of rat monocytes during acute kidney allograft rejection: increased expression of NKR-P1 and reduction of CD43. Scand J Immunol. 1998; 47(4):332-342. (Biology). View Reference
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Thomas ML, Green JR. Molecular nature of the W3/25 and MRC OX-8 marker antigens for rat T lymphocytes: comparisons with mouse and human antigens. Eur J Immunol. 1983; 13(10):855-858. (Clone-specific: Immunoprecipitation). View Reference
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Torres-Nagel N, Kraus E, Brown MH, et al. Differential thymus dependence of rat CD8 isoform expression.. Eur J Immunol. 1992; 22(11):2841-2848. (Clone-specific: Blocking, Immunoprecipitation, Western blot). View Reference
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Wallgren AC, Karlsson-Parra A, Korsgren O. The main infiltrating cell in xenograft rejection is a CD4+ macrophage and not a T lymphocyte. Transplantation. 1995; 60(6):594-601. (Clone-specific: Immunohistochemistry). View Reference
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.