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      RB705 Mouse Anti-Human CD3
      Product Details
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      BD OptiBuild™
      CD3E; CD3e; T-cell surface antigen T3/Leu-4 epsilon; T3E; TCRE
      Human (Tested in Development)
      Mouse BALB/c x A/J, also known as CAF1 IgG2a, κ
      Sheep Erythrocyte Rosette-purified Human T Cells
      Flow cytometry (Qualified)
      0.2 mg/ml
      VI 6T-R3, 6T-CD3.3
      916
      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 to the dye under optimum conditions that minimize unconjugated dye and antibody.
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      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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      3. For U.S. patents that may apply, see bd.com/patents.
      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      6. 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.
      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. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      10. An isotype control should be used at the same concentration as the antibody of interest.
      11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      12. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
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      757705 Rev. 1
      Antibody Details
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      OKT3

      The OKT3 monoclonal antibody specifically recognizes the CD3 epsilon subunit (CD3e/CD3ε) of the CD3 complex which consists of four transmembrane proteins (γ, δ, ε, ζ) that are associated with the T cell antigen receptor (TCR) to form the CD3/TCR complex. The CD3 complex associates with either TCR αβ or TCR γδ heterodimers that are alternatively expressed by some thymocytes, T cells or NKT cells. The CD3 complex is required for the cell surface expression and signal-transducing functions of the TCR. The CD3 complex is expressed by ~60-85% thymocytes and by all peripheral mature T cells. CD3e is also known as T3E or TCRE. CD3e is a ~20 kDa unglycosylated type I transmembrane protein that is encoded by CD3E which belongs to the immunoglobulin superfamily (IgSF). CD3e has an Ig-like extracellular domain (ECD) and an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The OKT3 antibody can reportedly fix complement, stimulate T cell proliferation and cytokine production, and block the binding of other human CD3e-specific antibodies including UCHT1 and SK7.

      757705 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      757705 Rev.1
      Citations & References
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      View product citations for antibody "757705" on CiteAb

      Development References (12)

      1. 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, Radioimmunoassay). View Reference
      2. Emmrich F. Selective stimulation of human CD4 and CD8 T-cells by crosslining the T-cell receptor with subset-specific differentiation antigens. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:203-206.
      3. Ernst DN, Shih CC. CD3 complex. J Biol Regul Homeost Agents. 2000; 14(3):226-229. (Biology). View Reference
      4. Horibe K, Knowles RW, Naito K, Morishima Y, Dupont B. Analysis of T lymphocyte antibody specificities: Comparison of serology with immunoprecipitation patterns. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies. Berlin New York: Springer-Verlag; 1984:212-224.
      5. Kung P, Goldstein G, Reinherz EL, Schlossman SF. Monoclonal antibodies defining distinctive human T cell surface antigens.. Science. 1979; 206(4416):347-9. (Immunogen: Cytotoxicity, Flow cytometry, Radioimmunoassay). View Reference
      6. Kurrle R, Seyfert W, Trautwein A, Seiler FR. T cell activation by CD3 antibodies. In: Reinherz EL. Ellis L. Reinherz .. et al., ed. Leukocyte typing II. New York: Springer-Verlag; 1986:137-146.
      7. Li B, Wang H, Dai J, et al. Construction and characterization of a humanized anti-human CD3 monoclonal antibody 12F6 with effective immunoregulation functions.. Immunology. 2005; 116(4):487-98. (Clone-specific: Blocking, Flow cytometry). View Reference
      8. Semnani R, Nutman TB, Corrado G, Hochman P, Shaw S, Van Seventer GA. Costimulation mediated by purified ICAM-1 and LFA-3 regulates differential stimulation and cytokine secretion of human 'naive' and 'memory' CD4+ T cells. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens. Oxford: Oxford University Press; 1995:1488-1491.
      9. Touraine JL, Favrot MC, Ansary ME, Cordier G, de bouteiller O. Phenotype of prothymocytes from human bone marrow determined by monoclonal antibodies: Modification induced by thymic factots. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies. Berlin New York: Springer-Verlag; 1984:298-311.
      10. Tunnacliffe A, Olsson C, Traunecker A, Krissansen GW, Karjalainen K, de la Hera A. The majority of CD3 epitopes are conferred by the epsilon chain. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:295-296.
      11. Van Wauwe JP, De Mey JR, Goossens JG. OKT3: a monoclonal anti-human T lymphocyte antibody with potent mitogenic properties.. J Immunol. 1980; 124(6):2708-13. (Clone-specific: Functional assay). View Reference
      12. 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: Functional assay). View Reference
      View All (12) View Less
      757705 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.