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Alexa Fluor® 700 Hamster anti-Mouse CD3e
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Alexa Fluor® 700 Hamster anti-Mouse CD3e

CD3e expression in spleen.  C57BL/6 splenocytes were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048 or 553049), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553032 or 553033), and either Alexa Fluor® 700 Hamster IgG2, λ Isotype Control (Cat. no. 557985, left panel) or Alexa Fluor® 700 Hamster anti-Mouse CD3e (right panel).  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Alexa Fluor® 700 Hamster anti-Mouse CD3e

CD3e expression in thymus.  C57BL/6 thymocytes were stained with either the same isotype control (dashed line) or Alexa Fluor® 700 Hamster anti-Mouse CD3e (solid line).  Flow cytometry was performed on a BD™ LSR II flow cytometry system

CD3e expression in spleen.  C57BL/6 splenocytes were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048 or 553049), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553032 or 553033), and either Alexa Fluor® 700 Hamster IgG2, λ Isotype Control (Cat. no. 557985, left panel) or Alexa Fluor® 700 Hamster anti-Mouse CD3e (right panel).  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

CD3e expression in thymus.  C57BL/6 thymocytes were stained with either the same isotype control (dashed line) or Alexa Fluor® 700 Hamster anti-Mouse CD3e (solid line).  Flow cytometry was performed on a BD™ LSR II 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)
0.2 mg/ml
12501
AB_396972
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 Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.

Recommended Assay Procedures

Note:  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.

Note:  Alexa Fluor® 700 conjugates of the hamster IgG2 isotype reportedly can non-specifically stain myeloid cells from mouse bone marrow.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. 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.
  4. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
557984 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.

557984 Rev. 1
Format Details
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Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
557984 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: Flow cytometry). 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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557984 Rev. 1

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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.