System Certification ISO 9001:2000
Exbio Antibodies CZ / SK eShop
by categories | a-z list | Application type: Flow Cytometry | Immunohistochemistry | Functional Studies

Mouse Monoclonal to ZAP-70

ZAP-03 (IgG1)

Technical Information Request    Inquiry for Bulk Prices

Product No. Form Quantity Stock Price Datasheet Shop
  11-488-C025 purified 0.025 mg yes choose region PDF datasheetHTML datasheet buy
  11-488-C100 purified 0.1 mg yes choose region PDF datasheetHTML datasheet buy
- In stock! Ready for immediate despatch - Manufacture on order

Background

The ZAP-70 (zeta-associated protein of 70 kDa) tyrosine kinase was identified as a tyrosine phosphoprotein that associates with TCR zeta subunit and undergoes tyrosine phosphorylation following TCR stimulation. ZAP-70 is a Syk family tyrosine kinase primarily expressed in T and NK cells that plays an essential role in signaling through the TCR. TCR-mediated activation of T cells is crucial to the immune response. In humans, ZAP-70 gene mutations resulting in lower ZAP-70 protein expression levels or expression of catalytically inactive ZAP-70 proteins, have been identified. ZAP-70 deficiency results in the absence of mature CD8+ T cells and the prevention of TCR-mediated activation of CD4+ T cells, and it can lead to severe combined immunodeficiency.
In patients with chronic lymphocytic leukemia (B-CLL), ZAP-70 expression on B cell was shown to be correlated with disease progression and survival. ZAP-70 contains two N-terminal SH2 domains (Src homology domain 2) and a C-terminal kinase domain. During T cell activation, the binding of ZAP-70 SH2 domains to the phosphorylated zeta subunit on the activated TCR complex causes a colocalization with the Lck tyrosine kinase that phosphorylates ZAP-70 on Tyr493 in the activation loop. ZAP-70 autophosphorylates multiple tyrosines in the region between the SH2 domains and the kinase domain, including the binding sites for additional SH2-containing signaling proteins such as SLP76, LAT, Lck, PLCgamma1, Vav, Shc, Ras-GAP, and Abl. ZAP-70-mediated activation of these downstream effectors leads to the release of intracellular calcium stores, and the transcription of interleukin-2 and other genes important for an immune response.

Specificity

The antibody ZAP-03 reacts with ZAP-70, a 70 kDa protein tyrosine kinase expressed in T and NK cells.
ZAP-70 is a molecule susceptible to degradation. It is recommended to use freshly prepared cell lysates (protease inhibitors are essential) to avoid non-specific staining of degradation products.

Regulatory Status

Immunogen

Bacterially expressed fusion protein representing C-terminal part (160 amino acids) of human ZAP-70 with histidine tag

Species Reactivity:

  • Human

Negative Species:

Applications:

  • Flow Cytometry
    Recommended dilution: 2-5 μg/ml
    Positive control:
    HPB-ALL human peripheral blood T cell leukemia cell line
    Application note: Intracellular staining
  • Western Blotting
    Recommended dilution: 0,5 μg/ml
    Positive control:
    HPB-ALL human peripheral blood T cell leukemia cell line
    Negative control:
    RAMOS human Burkitt lymphoma cell line
    Sample preparation:
    Resuspend approx. 50 mil. cells in 1 ml cold lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate (1:1) with reducing Laemmli SDS-PAGE sample buffer. Boil for 5 min.
    Application note: Reducing conditions. SDS-PAGE (10% separating gel).
  • Immunocytochemistry
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

  • *Ishaq M, DeGray G, Natarajan V: Evidence for the involvement of tyrosine kinase ZAP 70 in nuclear retinoid receptor-dependent transactivation in T lymphocytes. J Biol Chem. 2005 Oct 7;280(40):34152-8. [Abstract] [Full Text]
  • *Schneider H, Smith X, Liu H, Bismuth G, Rudd CE: CTLA-4 disrupts ZAP70 microcluster formation with reduced T cell/APC dwell times and calcium mobilization. Eur J Immunol. 2008 Jan;38(1):40-7. [Abstract]
  • *Chan AC, Irving BA, Fraser JD, Weiss A: The zeta chain is associated with a tyrosine kinase and upon T-cell antigen receptor stimulation associates with ZAP-70, a 70-kDa tyrosine phosphoprotein. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9166-70. [Abstract] [Full Text]
  • Product Specific References

  • *Hrdinka M, Dráber P, Stepánek O, Ormsby T, Otáhal P, Angelisová P, Brdicka T, Paces J, Horejsí V, Drbal K: PRR7 is a transmembrane adaptor protein expressed in activated T cells involved in regulation of T cell receptor signaling and apoptosis. J Biol Chem. 2011 Jun 3;286(22):19617-29. [Abstract] [Full Text]
  • For research use only. Not for drug, diagnostic or other use.

    Related Products

  • Mouse IgG1 Isotype Control
  • Example Data


    ZAP-03
    Fig. 1.

    Fig. 1. Western Blotting analysis (reducing conditions) of HPB-ALL peripheral blood T cell leukemia cell line.
    Lane 1, 2, 3, 4: immunostaining with dilution range of anti-ZAP-70 (ZAP-03; 4 μg/ml (1), 2 μg/ml (2), 1 μg/ml (3), 0.5 μg/ml (4)
    Lane 5: immunostaining with anti-ZAP-70 comparative antibody
    Lane 6:
    immunostaining with Isotype mouse IgG1 control (PPV-06; cat. no. 11-457-C100)



    ZAP-03
    Fig. 2.

    Fig. 2. Western Blotting analysis (reducing conditions) of HPB-ALL peripheral blood T cell leukemia cell line (1, 3) and RAMOS human Burkitt lymphoma cell line (2, 4);
    Lane 1, 2:
    immunostaining with anti-ZAP-70 (ZAP-03; 0.5 μg/ml)
    Lane 3, 4: immunostaining with Isotype mouse IgG1 control (PPV-06; cat. no. 11-457-C100)






    Unless indicated otherwise, all products are For Research Use Only and not for diagnostic or therapeutic use. Not for resale or transfer either as a stand-alone product or as a component of another product without written consent of EXBIO. EXBIO will not be held responsible for patent infringement or other violations that may occur with the use of our products. All orders are accepted subject to EXBIO´s term and conditions which are available at www.exbio.cz.
    © 2003 EXBIO. All rights reserved.
    terms and conditions | privacy policy