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Alexa Fluor® 647 Mouse Anti-Human CD3 ζ (CD247)
Alexa Fluor® 647 Mouse Anti-Human CD3 ζ (CD247)

Two-color flow cytometric analysis of CD3ζ (CD247) expression on human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were fixed and permeabilized with the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 (CD3ε; Cat. No. 562426/562427) and with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD3ζ (CD247) antibody at 1 µg/test (Cat. No. 566651). Flow-cytometric pseudocolor dot plots showing the correlated expression of CD3ε versus CD3ζ (CD247) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact human peripheral blood leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of CD3ζ (CD247) expression on human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were fixed and permeabilized with the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 (CD3ε; Cat. No. 562426/562427) and with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD3ζ (CD247) antibody at 1 µg/test (Cat. No. 566651). Flow-cytometric pseudocolor dot plots showing the correlated expression of CD3ε versus CD3ζ (CD247) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact human peripheral blood leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
CD247; CD3-ZETA; CD3Z; TCR zeta chain; TCRZ; T3Z
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human TCR zeta Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
AB_2869808
Aqueous buffered solution containing protein stabilizer and ≤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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566651 Rev. 1
Antibody Details
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6B10.2

The 6B10.2 monoclonal antibody specifically recognizes CD3 zeta (CD3Z or CD3ζ), which is also known as either CD247 or the invariant TCR zeta chain (TCRZ, T3Z, or TCRζ). CD3ζ (CD247) is a ~16 kDa single-pass, type I transmembrane protein that belongs to the CD3Z/FCER1G family. CD3ζ (CD247) has a short extracellular domain of nine amino acids and a cytoplasmic domain that contains three immunoreceptor tyrosine-based activation motifs (ITAMs).  Disulfide-linked CD3ζ homodimers usually associate with other transmembrane CD3 protein subunits (CD3δ, CD3ε, and CD3γ) to form the CD3 signaling complex which is linked to the clonotypic T cell receptor (TCR) for antigen, αβ TCR or γδ TCR. CD3ζ (CD247) is also expressed by natural killer (NK) cells where it can function as a signaling subunit associated with CD16a (FcγRIIIa), a receptor for IgG complexes. Human CD16a can couple with either CD3ζ or FcεRIγ homodimers, or CD3ζ-FcεRIγ heterodimers that serve to transduce IgG-bound CD16a-mediated signals intracellularly. Low or absent CD3ζ (CD247) expression is reportedly associated with impaired immune responses leading to some chronic infections or autoimmune diseases, and certain cancers. The 6B10.2 antibody is directed against the cytoplasmic domain of human CD3ζ (CD247).

        

566651 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566651 Rev.1
Citations & References
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Development References (6)

  1. Lanier LL. Up on the tightrope: natural killer cell activation and inhibition.. Nat Immunol. 2008; 9(5):495-502. (Biology). View Reference
  2. Myers MD, Sosinowski T, Dragone LL, et al. Src-like adaptor protein regulates TCR expression on thymocytes by linking the ubiquitin ligase c-Cbl to the TCR complex.. Nat Immunol. 2006; 7(1):57-66. (Clone-specific: Flow cytometry). View Reference
  3. Nambiar MP, Fisher CU, Warke VG, et al. Reconstitution of deficient T cell receptor zeta chain restores T cell signaling and augments T cell receptor/CD3-induced interleukin-2 production in patients with systemic lupus erythematosus.. Arthritis Rheum. 2003; 48(7):1948-55. (Clone-specific: Flow cytometry). View Reference
  4. Reichert TE, Day R, Wagner EM, Whiteside TL. Absent or low expression of the zeta chain in T cells at the tumor site correlates with poor survival in patients with oral carcinoma.. Cancer Res. 1998; 58(23):5344-7. (Clone-specific: Immunohistochemistry). View Reference
  5. Witherden D, van Oers N, Waltzinger C, Weiss A, Benoist C, Mathis D. Tetracycline-controllable selection of CD4(+) T cells: half-life and survival signals in the absence of major histocompatibility complex class II molecules.. J Exp Med. 2000; 191(2):355-64. (Clone-specific: Immunoprecipitation). View Reference
  6. van Oers NS, von Boehmer H, Weiss A. The pre-T cell receptor (TCR) complex is functionally coupled to the TCR-zeta subunit.. J Exp Med. 1995; 182(5):1585-90. (Immunogen: Immunoprecipitation). View Reference
View All (6) View Less
566651 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.