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BUV395 Hamster Anti-Mouse CD3e
BUV395 Hamster Anti-Mouse CD3e

Two-color flow cytometric analysis of CD3e expression on mouse splenocytes. BALB/c splenic leucocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and APC Rat Anti-Mouse CD8a (Cat. No. 553035/561093) antibodies and either BD Horizon™ BUV395 Armenian Hamster IgG1, κ Isotype Control (Cat. No. 563559; Left Panel) or BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565/565992; Right Panel). The two-color dot plots show CD3 (or Ig Isotype Control staining) versus CD4 and CD8 derived from events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometry System.

Two-color flow cytometric analysis of CD3e expression on mouse splenocytes. BALB/c splenic leucocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and APC Rat Anti-Mouse CD8a (Cat. No. 553035/561093) antibodies and either BD Horizon™ BUV395 Armenian Hamster IgG1, κ Isotype Control (Cat. No. 563559; Left Panel) or BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565/565992; Right Panel). The two-color dot plots show CD3 (or Ig Isotype Control staining) versus CD4 and CD8 derived from events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometry System.

Product Details
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BD Horizon™
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)
0.2 mg/ml
12501
AB_2738278
Aqueous buffered solution containing BSA 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 with BD Horizon BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the Brilliant Stain Buffer (Cat. No. 563794/566349).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563565 Rev. 4
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.

The antibody was conjugated to BD Horizon™ BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm  and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon™ BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

563565 Rev. 4
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
563565 Rev.4
Citations & References
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Development References (14)

  1. Castro JE, Listman JA, Jacobson BA, et al. Fas modulation of apoptosis during negative selection of thymocytes. Immunity. 1996; 5(6):617-627. (Clone-specific: Activation, Apoptosis). View Reference
  2. 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.
  3. Ernst DN, Weigle WO, McQuitty DN, Rothermel AL, Hobbs MV. Stimulation of murine T cell subsets with anti-CD3 antibody. Age-related defects in the expression of early activation molecules. J Immunol. 1989; 142(5):1413-1421. (Clone-specific: Activation, Functional assay, Stimulation). View Reference
  4. 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
  5. 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
  6. 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. (Clone-specific: Activation, Flow cytometry, Immunoprecipitation, Stimulation). View Reference
  7. 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, Flow cytometry, Immunoprecipitation, Inhibition, Stimulation). View Reference
  8. 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. (Clone-specific: Functional assay, Stimulation). View Reference
  9. 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. (Clone-specific: Blocking, Cytotoxicity, Immunoprecipitation, Radioimmunoassay). View Reference
  10. Radvanyi LG, Mills GB, Miller RG. Religation of the T cell receptor after primary activation of mature T cells inhibits proliferation and induces apoptotic cell death. J Immunol. 1993; 150(12):5704-5715. (Clone-specific: Activation, Apoptosis). View Reference
  11. 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. (Clone-specific: Activation, Calcium Flux, Flow cytometry, Western blot). View Reference
  12. 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
  13. Ucker DS, Meyers J, Obermiller PS. Activation-driven T cell death. II. Quantitative differences alone distinguish stimuli triggering nontransformed T cell proliferation or death. J Immunol. 1992; 149(5):1583-1592. (Clone-specific: Activation, Apoptosis). View Reference
  14. Wang R, Murphy KM, Loh DY, Weaver C, Russell JH. Differential activation of antigen-stimulated suicide and cytokine production pathways in CD4+ T cells is regulated by the antigen-presenting cell. J Immunol. 1993; 150(9):3832-3842. (Clone-specific: Activation, Apoptosis). View Reference
View All (14) View Less
563565 Rev. 4

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