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BUV661 Mouse Anti-Human CD3
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This product is the replacement for [565065].
BUV661 Mouse Anti-Human CD3

Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with either BD Horizon™ BUV661 Mouse IgG1, κ Isotype Control (Cat. No. 565064; dashed line histogram) or BD Horizon BUV661 Mouse Anti-Human CD3 antibody (Cat. No. 565065/565066; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was 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 either BD Horizon™ BUV661 Mouse IgG1, κ Isotype Control (Cat. No. 565064; dashed line histogram) or BD Horizon BUV661 Mouse Anti-Human CD3 antibody (Cat. No. 565065/565066; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was 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_2869644
Aqueous buffered solution containing ≤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 BUV661 under optimum conditions, and unconjugated antibody and free BD Horizon BUV661 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. An isotype control should be used at the same concentration as the antibody of interest.
  3. BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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 BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/30-nm filter).

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Catalog number 563794).

Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.

Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 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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BUV661
Ultraviolet 355 nm
350 nm
660 nm
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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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.