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Alexa Fluor® 647 Mouse Anti-Rat CD8a
Alexa Fluor® 647 Mouse Anti-Rat CD8a

Flow cytometric analysis of CD8a expression on rat splenocytes. Splenocytes from a Lewis rat were stained with the Alexa Fluor® 647 Mouse Anti-Rat CD8a antibody (Cat. No. 561611) in conjunction with a PE Mouse Anti-Rat CD3 antibody (Cat. No. 554833). The two-color flow cytometric dot plot showing the correlated expression of CD3 versus CD8a was derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD8a expression on rat splenocytes. Splenocytes from a Lewis rat were stained with the Alexa Fluor® 647 Mouse Anti-Rat CD8a antibody (Cat. No. 561611) in conjunction with a PE Mouse Anti-Rat CD3 antibody (Cat. No. 554833). The two-color flow cytometric dot plot showing the correlated expression of CD3 versus CD8a was derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
Cd8a; CD8α; CD8 alpha; OX-8 membrane antigen
Rat (QC Testing)
Mouse BALB/c IgG1, κ
High-molecular-weight rat thymocyte glycoproteins
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_10893006
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
561611 Rev. 1
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.

561611 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
561611 Rev.1
Citations & References
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Development References (18)

  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. 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
  12. 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
  13. 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
  14. Shapiro HM. Practical Flow Cytometry, 3rd Edition. New York: Wiley-Liss, Inc; 1995:280-281.
  15. 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
  16. 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). View Reference
  17. 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
  18. 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 (18) View Less
561611 Rev. 1

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