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Biotin Hamster Anti-Mouse CD3e
Biotin Hamster Anti-Mouse CD3e

CD3e expression in spleen and thymus. BALB/c splenocytes were simultaneously stained with PE-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553048/553049, Panels A and C), PE-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553032/553033, left panels), and biotinylated mAb 500A2 (bottom left panel), followed by Avidin-FITC (Cat. No. 554057, left panels). BALB/c thymocytes were stained with biotinylated 500A2 (bottom right panel) followed by Avidin-FITC (right panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.

CD3e expression in spleen and thymus. BALB/c splenocytes were simultaneously stained with PE-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553048/553049, Panels A and C), PE-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553032/553033, left panels), and biotinylated mAb 500A2 (bottom left panel), followed by Avidin-FITC (Cat. No. 554057, left panels). BALB/c thymocytes were stained with biotinylated 500A2 (bottom right panel) followed by Avidin-FITC (right panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Product Details
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BD Pharmingen™
CD3; CD3 epsilon; Cd3e; CD3ε; T3e
Mouse (QC Testing)
Syrian Hamster IgG2, κ
Mouse T-cell receptor
Flow cytometry (Routinely Tested)
0.5 mg/ml
12501
AB_394728
Aqueous buffered solution containing ≤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 biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

It has been observed that pre-incubation of thymus cell suspensions at 37°C for 2 to 4 hours prior to staining enhances the ability of anti-CD3e and anti-αβ TCR mAbs to detect the T-cell receptor on immature thymocytes.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
553239 Rev. 12
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.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

553239 Rev. 12
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
553239 Rev.12
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. (Biology). 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. (Biology). 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. (Methodology). View Reference
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553239 Rev. 12

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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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.