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Purified Hamster Anti-Mouse CD3e
Purified 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, left panels), PE-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553032/553033, left panels), and purified mAb 500A2 (bottom left panel), followed by FITC-conjugated anti-hamster IgG mAb G192-1 (Cat. No. 554026, left panels). BALB/c thymocytes were stained with purified 500A2 (bottom right panel) followed by FITC-conjugated G192-1 mAb (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, left panels), PE-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553032/553033, left panels), and purified mAb 500A2 (bottom left panel), followed by FITC-conjugated anti-hamster IgG mAb G192-1 (Cat. No. 554026, left panels). BALB/c thymocytes were stained with purified 500A2 (bottom right panel) followed by FITC-conjugated G192-1 mAb (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), Immunohistochemistry-frozen (Tested During Development), (Co)-stimulation, Immunoprecipitation (Reported)
0.5 mg/ml
12501
AB_394727
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. Store undiluted at 4°C.

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. For IHC, we recommend the use of purified 500A2 mAb in our special formulation for immunohistochemistry, Cat. No. 550277. For activation of mouse T lymphocytes by plate-bound anti-CD3e mAb, we recommend BD Biosciences Biocoat™ 96-Well Clear Anti-Mouse CD3 T-Cell Activation Plates, Cat. No. 354720.

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. 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.
553238 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.

553238 Rev. 12
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
553238 Rev.12
Citations & References
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View product citations for antibody "553238" on CiteAb

Development References (6)

  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: (Co)-stimulation). View Reference
  3. Jacobs H, Vandeputte D, Tolkamp L, de Vries E, Borst J, Berns A. CD3 components at the surface of pro-T cells can mediate pre-T cell development in vivo. Eur J Immunol. 1994; 24(4):934-939. (Clone-specific: (Co)-stimulation). View Reference
  4. 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
  5. 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: (Co)-stimulation). View Reference
  6. 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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553238 Rev. 12

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.