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CD3e expression in spleen and thymus. BALB/c splenocytes were simultaneously stained with PE-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553049), PE-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553033) and either purified anti-mouse CD3e mAb 145-2C11 (bottom left panel) or isotype control (top left panel), followed by staining with a FITC-conjugated anti-hamster IgG mAb cocktail (Cat. No. 554011). BALB/c thymocytes were also stained with purified anti-mouse CD3e 145-2C11 (bottom right panel) or isotype control (top right panel) followed by staining with a FITC-conjugated anti-hamster IgG mAb cocktail. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
BD Pharmingen™ Purified Hamster Anti-Mouse CD3e
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
Product Notices
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
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 (13)
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Chai JG, Lechler RI. Immobilized anti-CD3 mAb induces anergy in murine naive and memory CD4+ T cells in vitro. Int Immunol. 1997; 9(7):935-944. (Biology: Activation, Stimulation). View Reference
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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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Payer E, Elbe A, Stingl G. Circulating CD3+/T cell receptor V gamma 3+ fetal murine thymocytes home to the skin and give rise to proliferating dendritic epidermal T cells. J Immunol. 1991; 146(8):2536-2543. (Biology: Immunofluorescence). 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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Salvadori S, Gansbacher B, Pizzimenti AM, Zier KS. Abnormal signal transduction by T cells of mice with parental tumors is not seen in mice bearing IL-2-secreting tumors. J Immunol. 1994; 153(11):5176-5182. (Biology: Western blot). 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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Vossen AC, Tibbe GJ, Kroos MJ, van de Winkel JG, Benner R, Savelkoul HF. Fc receptor binding of anti-CD3 monoclonal antibodies is not essential for immunosuppression, but triggers cytokine-related side effects. Eur J Immunol. 1995; 25(6):1492-1496. (Biology: Blocking). View Reference
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Wagner DH Jr, Hagman J, Linsley PS, Hodsdon W, Freed JH, Newell MK. Rescue of thymocytes from glucocorticoid-induced cell death mediated by CD28/CTLA-4 costimulatory interactions with B7-1/B7-2. J Exp Med. 1996; 184(5):1631-1638. (Biology: Cytotoxicity). View Reference
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