CD3 expression and function Left Panel -Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes (PBL). Peripheral blood was incubated with either Purified Mouse IgG2a κ Isotype Control (Cat. No. 553454; dashed line histogram) or Purified NA/LE Mouse Anti-Human CD3 antibody (Cat. No. 566685/567107/567118; solid line histogram). The cells were washed and stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Right Panel - Multicolor flow cytometric analysis of CD25 expression on unstimulated and Anti-CD3 plus Anti-CD28 antibody-stimulated human PBL. Human peripheral blood mononuclear cells (PBMC) were not stimulated [Unstimulated: Bottom red-colored histograms] or were stimulated (72 h) with either soluble Purified NA/LE Mouse Anti-Human CD3 antibody (0.5 μg OKT-3 Ab/ml) plus soluble Purified NA/LE Mouse Anti-Human CD28 antibody (5 μg Ab/ml; Cat. No. 555725) [Midlevel orange histograms; sol CD3/sol CD28] or plate-bound Mouse Anti-Human CD3 antibody (5 μg OKT-3 Ab/ml overnight coating) plus Purified NA/LE Mouse Anti-Human CD28 antibody (5 μg Ab/ml) [Top blue histograms; pb CD3/sol CD28] as indicated. The cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD25 (Cat. No. 562442/564033), APC-H7 Mouse Anti Human CD4 (Cat. No. 560158/560251) and BD Horizon™ BUV395 Mouse Anti-Human CD8 (Cat. No. 563795/ 563796) antibodies. The fluorescence histograms showing CD25 expression were derived from either CD4+ (Left Histograms) or CD8+ (Right Histograms) gated events with the light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
CD3 expression and function Left Panel -Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes (PBL). Peripheral blood was incubated with either Purified Mouse IgG2a κ Isotype Control (Cat. No. 553454; dashed line histogram) or Purified NA/LE Mouse Anti-Human CD3 antibody (Cat. No. 566685/567107/567118; solid line histogram). The cells were washed and stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Right Panel - Multicolor flow cytometric analysis of CD25 expression on unstimulated and Anti-CD3 plus Anti-CD28 antibody-stimulated human PBL. Human peripheral blood mononuclear cells (PBMC) were not stimulated [Unstimulated: Bottom red-colored histograms] or were stimulated (72 h) with either soluble Purified NA/LE Mouse Anti-Human CD3 antibody (0.5 μg OKT-3 Ab/ml) plus soluble Purified NA/LE Mouse Anti-Human CD28 antibody (5 μg Ab/ml; Cat. No. 555725) [Midlevel orange histograms; sol CD3/sol CD28] or plate-bound Mouse Anti-Human CD3 antibody (5 μg OKT-3 Ab/ml overnight coating) plus Purified NA/LE Mouse Anti-Human CD28 antibody (5 μg Ab/ml) [Top blue histograms; pb CD3/sol CD28] as indicated. The cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD25 (Cat. No. 562442/564033), APC-H7 Mouse Anti Human CD4 (Cat. No. 560158/560251) and BD Horizon™ BUV395 Mouse Anti-Human CD8 (Cat. No. 563795/ 563796) antibodies. The fluorescence histograms showing CD25 expression were derived from either CD4+ (Left Histograms) or CD8+ (Right Histograms) gated events with the light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
BD Pharmingen™
CD3E; CD3e; T-cell surface antigen T3/Leu-4 epsilon; T3E; TCRE
Human (QC Testing)
Mouse BALB/c x A/J, also known as CAF1 IgG2a, κ
Sheep Erythrocyte Rosette-purified Human T Cells
Flow cytometry (Routinely Tested), Activation (Tested During Development)
1.0 mg/ml
VI 6T-R3, 6T-CD3.3
No azide/low endotoxin: Aqueous buffered solution containing no preservative, sterile filtered(0.2µm pore size membrane). Endotoxin level is ≤0.1 EU/µg (≤0.01 ng/µg) of protein as determined by the LAL assay.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Recommended Assay Procedures
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 (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Brilliant Stain Buffer Plus RUO
Size
1000 Tests
Cat No.
566385
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Purified Mouse IgG2a κ Isotype Control RUO
Size
0.5 mg
Cat No.
553454
567118 Rev. 2
Antibody Details
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.
567118 Rev. 2
Format Details
NA/LE
NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
NA/LE
567118 Rev.2
Citations & References
Development References (12)
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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
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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.
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Ernst DN, Shih CC. CD3 complex. J Biol Regul Homeost Agents. 2000; 14(3):226-229. (Biology). View Reference
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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.
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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
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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.
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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
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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.
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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.
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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.
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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
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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
567118 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.
