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Mouse Monoclonal to CD3 zeta (Phospho-Tyr72)

EM-26 (IgG1)

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Background

CD3 complex is crucial in transducing antigen-recognition signals into the cytoplasm of T cells and in regulating the cell surface expression of the TCR complex. T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta (CD247). These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR-induced growth arrest, cell survival and proliferation.

Specificity

The mouse monoclonal antibody EM-26 recognizes phosphorylated tyrosine 72 of CD3 zeta chain (CD247), which is a component of TCR/CD3 complex expressed on T cells.

Regulatory Status

Immunogen

A phospho specific peptide corresponding to the amino acids surrounding tyrosine 72 of mouse CD3 zeta linked to KLH

Species Reactivity:

  • Human
  • Mouse

Negative Species:

Applications:

  • Flow Cytometry
    Recommended dilution: 2 - 10 μg/ml
    Positive control: Jurkat cells treated with pervanadate; T-cells from lymph nodes of OT-1 mouse treated with pervanadate
  • Western Blotting
    Recommended dilution: 2 - 5 μg/ml
    Positive control: Jurkat cells lysate treated with pervanadate; Splenocyte lysate of Balb/c or F1 mouse treated with pervanadate
    Application note: Non-reducing conditions recommended
  • Immunocytochemistry
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

Product Specific References

  • *Dopfer EP, Schöpf B, Louis-Dit-Sully C, Dengler E, Höhne K, Klescová A, Prouza M, Suchanek M, Reth M, Schamel WW: Analysis of novel phospho-ITAM specific antibodies in a S2 reconstitution system for TCR-CD3 signalling. Immunol Lett. 2010 May 4;130(1-2):43-50 [Abstract]
  • *Borroto A, Arellano I, Dopfer EP, Prouza M, Suchànek M, Fuentes M, Orfao A, Schamel WW, Alarcón B: Nck recruitment to the TCR required for ZAP70 activation during thymic development. J Immunol. 2013 Feb 1;190(3):1103-12. [Abstract]
  • For research use only. Not for drug, diagnostic or other use.

    Example Data

    WB on mutants
    Fig. 1.


    Fig. 1. Reactivity of the monoclonal antibodies EM-26 (anti-CD3 zeta phospho-Tyr72) and EM-17 (anti-CD3 zeta phospho-Tyr153) with phosphorylated particular human CD3 zeta mutants. The Y1F and Y6F mutatants lack phosphotyrosine 72 and 153, respectively.


    synapse EM-26
    Fig. 2.



    Fig. 2. Detection of phosphorylated CD3 zeta (EM-26; light blue) in immunological synapse formed between the lymph node naïve T cells from AND TCR transgenic mice and DCEK cells loaded with MCC peptide, after 20 min. Total CD3 zeta indicated in green, actin cytoskeleton in red. 

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