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BUV496 Mouse Anti-Human CD3
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BUV496 Mouse Anti-Human CD3
Multiparameter flow cytometric analysis using BD OptiBuild™ BUV496 Mouse Anti-Human CD3 antibody (Cat. No. 741133) on Human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.
Multiparameter flow cytometric analysis using BD OptiBuild™ BUV496 Mouse Anti-Human CD3 antibody (Cat. No. 741133) on Human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.
Product Details
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BD OptiBuild™
CD3E; T3E; TCRE; cd 3; cd-3; cd3; CD3-epsilon; 916
Human (Tested in Development)
Mouse IgG2a, κ
Flow cytometry (Qualified)
0.2 mg/ml
V 5T-CD03.05
916
AB_2870713
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 to the dye under optimum conditions that minimize unconjugated dye and antibody. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For U.S. patents that may apply, see bd.com/patents.
741133 Rev. 2
Antibody Details
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HIT3a

The HIT3a monoclonal antibody specifically binds to the human CD3ε-chain, a 20 kDa subunit of the CD3/T cell antigen receptor complex found on 70-80% of normal human peripheral blood lymphocytes and 60-85% of thymocytes. Studies from the HLDA Workshop show that this antibody can be mitogenic for T lymphocytes. The CD3 complex plays a role in signal transduction during antigen recognition by the T cell receptor. HIT3a antibody does not stain intracellular CD3 unlike the other CD3-specific clone, UCHT1 (Cat. No. 555330/550368).

741133 Rev. 2
Format Details
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BUV496
The BD Horizon Brilliant™ Ultraviolet 496 (BUV496) 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 496-nm. BUV496, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 500-nm (e.g., 515/30-nm bandpass filter). The acceptor dye can be excited by the Violet (405-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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BUV496
Ultraviolet 355 nm
350 nm
496 nm
741133 Rev.2
Citations & References
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Development References (4)

  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. (Biology). View Reference
  2. Boumsell L. T cell antigens: section report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:241-279.
  3. Ernst DN, Shih CC. CD3 complex. J Biol Regul Homeost Agents. 2000; 14(3):226-229. (Biology). View Reference
  4. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
View All (4) View Less
741133 Rev. 2

Please refer to Support Documents for Quality Certificates


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.