Skip to main content Skip to navigation
Hamburger Menu
Search icon
Location Selector Location Selector
Portugal (English)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Link Arrow
Solutions

Link Arrow
Discover & Learn

Link Arrow
Resources & Tools

Link Arrow
Support

Link Arrow
Close Menu
Website Feedback
Alert Icon
566696_566697_2.png

Multiparameter flow cytometric analysis of CD3 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, k Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 566696/566697; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

566697Image1.png

Multiparameter flow cytometric analysis of CD3 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, k Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 566696/566697; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

566696_566697_2.png 566697Image1.png Blue-Cap-Violet-Label.png

Multiparameter flow cytometric analysis of CD3 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, k Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 566696/566697; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

Multiparameter flow cytometric analysis of CD3 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, k Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 566696/566697; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

Product Details
Down Arrow Up Arrow


BD Horizon™
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)
5 µl
VI 6T-R3, 6T-CD3.3
916
AB_2869824
Aqueous buffered solution containing BSA and ≤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 and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Show More blue down arrow Show Less blue up arrow

Recommended Assay Procedures

Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended 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).

Show More blue down arrow Show Less blue up arrow

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  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. For U.S. patents that may apply, see bd.com/patents.
Show More blue down arrow Show Less blue up arrow
566696 Rev. 3
Antibody Details
Down Arrow Up Arrow
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.

566696 Rev. 3
Format Details
Down Arrow Up Arrow
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
566696 Rev.3
Citations & References
Down Arrow Up Arrow
View product citations for antibody "566696" 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
566696 Rev. 3

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.