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BV650 Mouse Anti-Human CD3
BV650 Mouse Anti-Human CD3
Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV650 Mouse Anti-Human CD3 antibody (Cat. No. 563851/563852; solid line histogram) or with BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometry System.
Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV650 Mouse Anti-Human CD3 antibody (Cat. No. 563851/563852; solid line histogram) or with BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometry System.
Product Details
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BD Horizon™
CD3e; CD3E; T3E; TCRE; T-cell surface antigen T3/Leu-4 epsilon
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human infant thymocytes and peripheral blood lymphocytes from a Sézary Syndrome donor
Flow cytometry (Routinely Tested)
5 µl
I WT3; III 471
916
AB_2744391
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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Brilliant Violet™ 650 is a trademark of Sirigen.
563852 Rev. 1
Antibody Details
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UCHT1

The UCHT1 monoclonal antibody specifically binds to the human CD3ε-chain, a 20-kDa subunit of the CD3/T cell antigen receptor complex. CD3ε is expressed on 70-80% of normal human peripheral blood lymphocytes and 60-85% of thymocytes. Studies from the HLDA Workshop show that this antibody is mitogenic for CD3ε-positive cells when used in conjunction with costimulatory agents such as pokeweed mitogen or anti-CD28 antibody. CD3 plays a central role in signal transduction during antigen recognition.  The UCHT1 antibody stains both surface and intracellular CD3ε unlike the other CD3 clone, HIT3a, that stains only extracellular CD3ε.

The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm.  BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter).  Due to the excitation and emission characteristics of the acceptor dye, there will be  spillover into the APC and Alexa Fluor® 700 detectors.  However, the spillover can be corrected through compensation as with any other dye combination.

563852 Rev. 1
Format Details
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BV650
The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV650
Violet 405 nm
406 nm
649 nm
563852 Rev.1
Citations & References
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Development References (6)

  1. Beverley PC, Callard RE. Distinctive functional characteristics of human "T" lymphocytes defined by E rosetting or a monoclonal anti-T cell antibody. Eur J Immunol. 1981; 11(4):329-334. (Immunogen: Flow cytometry, Fluorescence activated cell sorting). View Reference
  2. Burns GF, Boyd AW, Beverley PC. Two monoclonal anti-human T lymphocyte antibodies have similar biologic effects and recognize the same cell surface antigen. J Immunol. 1982; 129(4):1451-1457. (Clone-specific: Blocking, Functional assay, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
  3. Ernst DN, Shih CC. CD3 complex. J Biol Regul Homeost Agents. 2000; 14(3):226-229. (Biology). View Reference
  4. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  5. McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
  6. Van Wauwe JP, Goossens JG, Beverley PC. Human T lymphocyte activation by monoclonal antibodies; OKT3, but not UCHT1, triggers mitogenesis via an interleukin 2-dependent mechanism. J Immunol. 1984; 133(1):129-132. (Clone-specific: Flow cytometry, Functional assay, Stimulation). View Reference
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563852 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.