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V450 Hamster anti-Mouse CD3e
V450 Hamster anti-Mouse CD3e

Flow cytometric analysis of CD3 expression on mouse splenocytes using BD Horizon™ V450 anti-mouse CD3e. BALB/c mouse spleen cells were stained simultaneously with BD Horizon™ V450-conjugated anti-mouse CD3e antibody and Alexa Fluor® 647 anti-mouse CD45R (Cat. No.557683; right panel) or FITC anti-mouse CD4 (Cat. No.553047 ; left panel) antibody. Two-color flow cytometric dot plots showing the correlated expression patterns of CD3e and CD45R  or CD3e and CD4  were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSRII System.

Flow cytometric analysis of CD3 expression on mouse splenocytes using BD Horizon™ V450 anti-mouse CD3e. BALB/c mouse spleen cells were stained simultaneously with BD Horizon™ V450-conjugated anti-mouse CD3e antibody and Alexa Fluor® 647 anti-mouse CD45R (Cat. No.557683; right panel) or FITC anti-mouse CD4 (Cat. No.553047 ; left panel) antibody. Two-color flow cytometric dot plots showing the correlated expression patterns of CD3e and CD45R  or CD3e and CD4  were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSRII System.

Product Details
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BD Horizon™
CD3; CD3 epsilon; Cd3e; CD3ε; T3e
Mouse (QC Testing)
Syrian Hamster IgG2, κ
Mouse T-cell receptor
Flow cytometry (Routinely Tested)
0.2 mg/ml
12501
AB_2034005
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
560801 Rev. 1
Antibody Details
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500A2

The 500A2 monoclonal antibody specifically binds to the 25-kDa ε chain of the T-cell receptor-associated CD3 complex expressed on mouse thymocytes, mature T lymphocytes, and NKT cells. Plate-bound and soluble forms of the 500A2 antibody can activate T cells in vitro. Activation of a mouse T-cell clone by the 500A2 antibody can be blocked by Fab fragments of the GK1.5 anti-CD4 antibody. This suggests that the 500A2 antibody may bind an epitope on CD3e close to a site at which CD4 associates with the T-cell receptor. The 500A2 antibody reportedly does not to crossreact with rat leukocytes.

The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

560801 Rev. 1
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
V450
Violet 405 nm
405 nm
450 nm
560801 Rev.1
Citations & References
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Development References (5)

  1. Allison JP, Havran WL, Poenie M, et al. Expression and function of CD3 on murine thymocytes. In: Kappler J, Davis M, ed. The T-Cell Receptor, UCLA Symposia, 73rd Edition. Los Angeles: 1988:33-45.
  2. Havran WL, Poenie M, Kimura J, Tsien R, Weiss A, Allison JP. Expression and function of the CD3-antigen receptor on murine CD4+8+ thymocytes. Nature. 1987; 330(6144):170-173. (Clone-specific). View Reference
  3. 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. (Methodology). View Reference
  4. Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998; 160(1):1158-1165. (Clone-specific). View Reference
  5. 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). View Reference
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560801 Rev. 1

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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.