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APC Hamster Anti-Mouse CD3e
APC Hamster Anti-Mouse CD3e

Flow cytometric analysis of CD3e expression in mouse spleen (left panels) and thymus (right panels). BALB/c splenocytes were simultaneously stained with PE Rat anti-Mouse CD4 (Cat. No. 553048), PE Rat anti-Mouse CD8a (Cat. No. 553032), and APC Hamster anti-Mouse CD3e (Cat. No. 561826/553066; bottom left panel). BALB/c thymocytes were also stained with APC Hamster anti-Mouse CD3e (bottom right panel) or unstained (top right panel). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Flow cytometric analysis of CD3e expression in mouse spleen (left panels) and thymus (right panels). BALB/c splenocytes were simultaneously stained with PE Rat anti-Mouse CD4 (Cat. No. 553048), PE Rat anti-Mouse CD8a (Cat. No. 553032), and APC Hamster anti-Mouse CD3e (Cat. No. 561826/553066; bottom left panel). BALB/c thymocytes were also stained with APC Hamster anti-Mouse CD3e (bottom right panel) or unstained (top right panel). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Pharmingen™
CD3; CD3 epsilon; Cd3e; CD3ε; T3e
Mouse (QC Testing)
Armenian Hamster IgG1, κ
H-2Kb specific cytotoxic T lymphocyte clone BM10-37
Flow cytometry (Routinely Tested), Intracellular staining (flow cytometry) (Reported)
0.2 mg/ml
12501
AB_398529
Aqueous buffered solution containing protein stabilizer and ≤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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
561826 Rev. 23
Antibody Details
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145-2C11

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.

561826 Rev. 23
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 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.
altImg
APC
Red 627-640 nm
651 nm
660 nm
561826 Rev.23
Citations & References
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Development References (9)

  1. 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.
  2. Hattori N, Kawamoto H, Katsura Y. Isolation of the most immature population of murine fetal thymocytes that includes progenitors capable of generating T, B, and myeloid cells. J Exp Med. 1996; 184(5):1901-1908. (Biology: Flow cytometry, IC/FCM Block). View Reference
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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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561826 Rev. 23

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