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    PerCP Mouse Anti-Rat CD8a
    PerCP Mouse Anti-Rat CD8a
    The expression of CD8a on rat splenocytes. Single-cell suspensions of Lewis splenocytes were stained with PerCP-conjugated mAb OX-8 and PE-conjugated anti-rat CD3 mAb G4.18 (Cat. No. 554833). Note that the CD8a+CD3- population represents NK cells. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
    PerCP Mouse Anti-Rat CD8a
    The expression of CD8a on rat splenocytes. Single-cell suspensions of Lewis splenocytes were stained with PerCP-conjugated mAb OX-8 and PE-conjugated anti-rat CD3 mAb G4.18 (Cat. No. 554833). Note that the CD8a+CD3- population represents NK cells. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
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    BD Pharmingen™
    Rat (QC Testing)
    Mouse BALB/c IgG1, κ
    High-molecular-weight rat thymocyte glycoproteins
    Flow cytometry (Routinely Tested)
    0.2 mg/ml
    AB_397133
    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 with PerCP under optimum conditions, and unconjugated antibody and free PerCP were removed. Storage of PerCP conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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    Recommended Assay Procedures

    For tandem conjugates incorporating PerCP (e.g., PerCP-Cy5.5), the excitation and emission properties of PerCP and the kinetics of energy exchange between the fluorochromes of the tandem dye may limit their effectiveness on high-speed and/or sorting flow cytometers.

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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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    4. PerCP is a photosynthetic accessory pigment from Glenodinium species of dinoflagellates, which is excited by the 488-nm light of an Argon ion laser and fluoresces at 675 nm. Therefore, PerCP-labelled antibodies can be used with FITC- and R-PE–labelled reagents in most single-laser flow cytometers with no significant spectral overlap of PerCP fluorescence with that of FITC or R-PE. PerCP has been reported to undergo significant photobleaching, the magnitude of which increases as laser power is increased or beam focus is narrowed. For third-color flow¬cytometric analysis using ≥25-mW laser power, we recommend PE-Cy5-, PE-Cy7–, or PerCP-Cy5.5-conjugated reagents.
    5. 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.
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    558824 Rev. 8
    Antibody Details
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    OX-8

    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.

    558824 Rev. 8
    Format Details
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    PerCP
    PerCP dye is part of the BD blue family of dyes. This dye is a fluorescent protein complex with an excitation maximum (Ex Max) of 481 nm and an emission maximum (Em Max) at 675 nm. PerCP is designed to be excited by the blue laser (488 nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405 nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    altImg
    PerCP
    Blue 488 nm
    481 nm
    675 nm
    558824 Rev.8
    Citations & References
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    Development References (19)

    1. Afar B, Merrill J, Clark EA. Detection of lymphocyte subsets using three-color/single-laser flow cytometry and the fluorescent dye peridinin chlorophyll-alpha protein. J Clin Immunol. 1991; 11(5):254-261. (Biology). View Reference
    2. 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
    3. 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
    4. 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
    5. 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
    6. Greimers R, Trebak M, Moutschen M, Jacobs N, Boniver J. Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets. Cytometry. 1996; 23(3):205-217. (Biology). View Reference
    7. 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
    8. 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
    9. 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
    10. 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
    11. 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
    12. 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
    13. 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
    14. 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
    15. Shapiro HM. Practical Flow Cytometry, 3rd Edition. New York: Wiley-Liss, Inc; 1995:280-281.
    16. 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
    17. 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
    18. Waggoner AS, Ernst LA, Chen CH, Rechtenwald DJ. PE-CY5. A new fluorescent antibody label for three-color flow cytometry with a single laser. Ann N Y Acad Sci. 1993; 677:185-193. (Biology). View Reference
    19. 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
    View All (19) View Less
    558824 Rev. 8

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    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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