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Flow cytometric analysis of CD3e expression in spleen and thymus. BALB/c splenocytes were simultaneously stained with PE Rat Anti-Mouse CD4 (Cat. No. 553048), PE-Rat Anti-Mouse CD8a (Cat. No. 553032) and Biotin Hamster Anti-Mouse CD3e (Cat. No. 553060/553059; bottom left panel), followed by Avidin FITC (Cat. No. 554057). BALB/c thymocytes were also stained with Biotin Hamster Anti-Mouse CD3e, followed by Avidin FITC (Cat. No. 553059/553060; bottom right panel). Fluorescence contour plots and histograms were derived from gated events based on the forward and side light-scattering of viable splenocytes and thymocytes. Flow cytometry was performed on a BD FACScan™.
BD Pharmingen™ Biotin Hamster Anti-Mouse CD3e
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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
Product Notices
- 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.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
- 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.
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Companion Products
The 145-2C11 monoclonal antibody specifically binds to the 25-kDa ε chain of the T-cell receptor-associated CD3 complex that is expressed on thymocytes, mature T lymphocytes, and NK-T cells. The cytoplasmic domain of CD3e participates in the signal transduction events that activate several cellular biochemical pathways as a result of antigen recognition. Soluble 145-2C11 antibody can activate either unprimed (naive) or primed (memory/preactivated) T cells in vivo or in vitro, in the presence of Fc receptor-bearing accessory cells. In contrast, plate-bound 145-2C11 can activate T cells in the absence of accessory cells. Soluble 145-2C11 antibody has been reported to induce re-directed lysis of Fc receptor-bearing target cells by CTL clones and can also block lysis of specific target cells by antigen-specific CTL's. Under some conditions, T-cell activation by 145-2C11 antibody has been reported to result in apoptotic cell death. The 145-2C11 antibody does not cross-react with rat leukocytes. Preincubation of thymus cell suspensions at 37°C for 2-4 hours prior to staining reportedly enhances the ability of anti-CD3ε and anti-αβ TCR mAbs to detect the T-cell receptor on immature thymocytes.
Development References (9)
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Duke RC, Cohen JJ, Boehme SA, et al. Morphological, biochemical, and flow cytometric assays of apoptosis. In: Coligan J, Kruisbeek AM, Margulies D, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: John Wiley and Sons; 1995:3.17.1-3.17.33.
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Isakov N, Wange RL, Burgess WH, Watts JD, Aebersold R, Samelson LE. ZAP-70 binding specificity to T cell receptor tyrosine-based activation motifs: the tandem SH2 domains of ZAP-70 bind distinct tyrosine-based activation motifs with varying affinity. J Exp Med. 1995; 181(1):375-380. (Biology: Immunoprecipitation). View Reference
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Kruisbeek AM, Shevach EM. Proliferative assays for T cell function. Curr Protoc Immunol. 2004; 3:3.12.1-3.12.14. (Methodology: Activation, Stimulation). View Reference
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Kubo RT, Born W, Kappler JW, Marrack P, Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989; 142(8):2736-2742. (Biology). View Reference
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Leo O, Foo M, Sachs DH, Samelson LE, Bluestone JA. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987; 84(5):1374-1378. (Immunogen: Activation, Blocking, Cytotoxicity, Immunoprecipitation, Stimulation). View Reference
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Nakano H, Yamazaki T, Miyatake S, Nozaki N, Kikuchi A, Saito T. Specific interaction of topoisomerase II beta and the CD3 epsilon chain of the T cell receptor complex. J Biol Chem. 1996; 271(11):6483-6489. (Biology: Immunoprecipitation). View Reference
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Portoles P, Rojo J, Golby A, et al . Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation. J Immunol. 1989; 142(12):4169-4175. (Biology: Activation, Immunoprecipitation, Stimulation). View Reference
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Shinkai Y, Alt FW. CD3 epsilon-mediated signals rescue the development of CD4+CD8+ thymocytes in RAG-2-/- mice in the absence of TCR beta chain expression. Int Immunol. 1994; 6(7):995-1001. (Biology: Activation, Stimulation). View Reference
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Zheng B, Han S, Kelsoe G. T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1. J Exp Med. 1996; 184(3):1083-1091. (Biology: Immunohistochemistry). View Reference
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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.