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Two-color flow cytometric analysis of CD3e expressed on mouse splenocytes. BALB/c splenocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051) and APC Rat Anti-Mouse CD8a (Cat. No. 561093/553035) and either BD Horizon™ BV421 Armenian Hamster IgG1, κ Isotype Control (Cat. No. 562601; Left Panel) or BD Horizon™ BV421 Hamster Anti-Mouse CD3e antibody (Cat. No. 562600; Right Panel). The two-color flow cytometric dot plots showing CD3 (or Ig Isotype Control staining) versus CD4 and CD8 were derived from events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
BD Pharmingen™ BV421 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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- 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.
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.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates..
Development References (11)
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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). 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). 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). 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). View Reference
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Takizawa F, Kinet JP, Adamczewski M. Binding of phycoerythrin and its conjugates to murine low affinity receptors for immunoglobulin G. J Immunol Methods. 1993; 162(2):269-272. (Biology). View Reference
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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
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van Vugt MJ, van den Herik-Oudijk IE, van de Winkle JG. Binding of PE-CY5 conjugates to the human high-affinity receptor for IgG (CD64). Blood. 1996; 88(6):2358-2361. (Biology). View Reference
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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.