ZAP70 Antibody (2F3.2) - Azide and BSA Free

Immunocytochemistry/Immunofluorescence: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Immunofluorescence Analysis of PFA-fixed Jurkat cells labeled with ZAP70 Antibody (2F3.2) followed by Goat anti-Mouse IgG-CF488 (Green). The nuclear counterstain is Nucspot (Red).

Immunohistochemistry-Paraffin: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Human Tonsil stained with ZAP70 Monoclonal Antibody (2F3.2).

Flow Cytometry: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Flow Cytometric Analysis of PFA-fixed Jurkat cells. ZAP70 Antibody (2F3.2) followed by goat anti-Mouse IgG-CF488 (Blue); Isotype Control (Red).

Simple Western: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Electropherogram image of the corresponding Simple Western lane. ZAP70 antibody was used at 10 ug/ml dilution of Jurkat and H. Tonsil lysates(s) respectively.

Simple Western: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Lane view shows a specific band for ZAP70 in Jurkat cell lysate using 50 ug/mL antibody dilution. Electropherogram image of corresponding Simple Western lane view at WES molecular weight of 70 kDa. Image from an internal validation.

Simple Western: ZAP70 Antibody (2F3.2) - Azide and BSA Free [NBP2-47774] - Simple Western lane view shows a specific band for ZAP70 in 0.2 mg/ml of Jurkat (left) and H. Tonsil (right) lysate(s). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: Simple Western, Flow, ICC/IF, IHC, CyTOF-ready, ICC/IF
• Clone: 2F3.2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1.0 mg/ml

ZAP70 Antibody (2F3.2) - Azide and BSA Free Summary

• Description: 1.0 mg/ml of antibody purified from Bioreactor Concentrate by Protein A/G. Prepared in 10mM PBS WITHOUT BSA & azide. Also available at 200 ug/ml WITH BSA & azide (NBP2-44502).
  
  Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.
• Immunogen: Recombinant ZAP-70 protein including residues 1-254 and encompassing SH2 domains of human ZAP70 (Uniprot: P43403)
• Localization: Cell Surface and Cytoplasmic
• Marker: Chronic Lymphocytic Leukemia Marker
• Specificity: ZAP70 is a 70kDa protein tyrosine kinase found in T-cells and natural killer cells. Control of this protein translation is via the IgVH gene. In Western blotting of whole cell lysates of normal peripheral blood mononuclear cells, the antibody labels a band corresponding to ZAP70. In Western blotting of whole cell lysates of CD19-positive Purified leukemia cells from patients with Ig-unmutated and Ig-mutated CLL, the antibody labels a band corresponding to ZAP70 in the Ig-unmutated CLL samples, whereas no band is observed in the Ig-mutated CLL samples. In Western blotting of cell lysates of Jurkat cells (T-lymphoblastic cell line), the antibody labels a band of 70kDa protein. In Western blotting of cell lysates of A431 cells (carcinoma cell line), no band is observed. ZAP70 protein is expressed in leukemic cells of approximately 25% of chronic lymphocytic leukemia (CLL) cases as well. Anti-ZAP70 expression is an excellent surrogate marker for the distinction between the Ig-mutated (anti-ZAP70 negative) and Ig-unmutated (anti-ZAP70 positive) CLL subtypes and can identify patient groups with divergent clinical courses. The anti-ZAP70 positive Ig-unmutated CLL cases have been shown to have a poorer prognosis.
• Isotype: IgG2a Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: ZAP70
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.5 - 1 ug/million cells in 0.1 ml
  • Immunocytochemistry/ Immunofluorescence
  • Immunofluorescence 0.5 - 1.0 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 0.5 - 1.0 ug/ml
  • Simple Western 10 ug/ml
• Application Notes: Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined.
  In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: antibody dilution of 10 ug/mL.
• Theoretical MW: 70 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -20 to -80C. Avoid freeze-thaw cycles.
• Buffer: 10 mM PBS
• Preservative: No Preservative
• Concentration: 1.0 mg/ml
• Purity: Protein A or G purified

Alternate Names for ZAP70 Antibody (2F3.2) - Azide and BSA Free

• 70 kDa zeta-associated protein
• EC 2.7.10
• EC 2.7.10.2
• mrtle
• mur
• SRK
• SRKFLJ17679
• STD
• STDFLJ17670
• Syk-related tyrosine kinase
• tyrosine-protein kinase ZAP-70
• TZK
• ZAP70
• ZAP-70
• zeta-chain (TCR) associated protein kinase (70 kD)
• zeta-chain (TCR) associated protein kinase 70kDa
• zeta-chain associated protein kinase, 70kD

Background

Zap-70, a Syk-family protein tyrosine kinase, plays a critical role in mediating T cell signal transduction in response to T cell receptor (TCR) activation (1). TCR-mediated activation of the Src-family kinases, Lck and Fyn, results in tyrosine phosphorylation of the TCR zeta and CD3 chains. These domains serve as targets for binding of ZAP-70 via its tandem SH2 domains. This binding correlates with activation of ZAP-70, a critical event in T cell activation (2). Following TCR engagement, ZAP-70 is phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a trans-phosphorylation by the Src-family tyrosine kinase, Lck. Phosphorylation of Tyr319 is required for full activation and increased positive downstream regulation by ZAP-70 (3).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: ZAP70