Anti-ZAP70 Alexa Fluor® 647

Anti-ZAP70 Alexa Fluor® 647

• Background: The ZAP70 (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. ZAP70 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, ZAP70 gene mutations resulting in lower ZAP70 protein expression levels or expression of catalytically inactive ZAP70 proteins, have been identified. ZAP70 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), ZAP70 expression on B cell was shown to be correlated with disease progression and survival. ZAP70 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 ZAP70 on Tyr493 in the activation loop. ZAP70 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. ZAP70-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.
• Specifications: The mouse monoclonal antibody 1E7.2 recognizes ZAP70, a 70 kDa protein tyrosine kinase expressed in T and NK cells (an intracellular antigen). ZAP70 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. The epitope is located within "interdomain B".
• CAS Number: 9007-83-4
• Certification: RUO
• Host: Mouse
• Species Reactivity: Human, Mouse
• Immunogen: A KLH-coupled peptide corresponding to amino acids 282-307 of human ZAP70
• Target Antigen: ZAP70
• Isotype: Igg1 κ
• Clone: 1E7.2
• Applications: Flow cytometry: The reagent is designed for analysis of human blood cells using 4 μl reagent / 100 μl of whole blood or 106 cells in a suspension. The content of a vial (0.4 mL) is sufficient for 100 tests. Intracellular staining.
• Validated Applications: WB, ICC, FC
• Format: Alexa Fluor® 647
• Buffer: Stabilizing phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
• References & Citations: *Rossi FM, Del Principe MI, Rossi D, Irno Consalvo M, Luciano F, Zucchetto A, Bulian P, Bomben R, Dal Bo M, Fangazio M, Benedetti D, Degan M, Gaidano G, Del Poeta G, Gattei V: Prognostic impact of ZAP-70 expression in chronic lymphocytic leukemia: mean fluorescence intensity T/B ratio versus percentage of positive cells. J Transl Med. 2010 Mar 88:23., URL: http://www.ncbi.nlm.nih.gov/pubmed/20211015 ,*Vroblova V, Vrbacky F, Hrudkova M, Jankovicova K, Schmitzova D, Maly J, Krejsek J, Smolej L: Significant change in ZAP-70 expression during the course of chronic lymphocytic leukemia. Eur J Haematol. 2010 Jun84(6):513-7., URL: https://www.ncbi.nlm.nih.gov/pubmed/20132303,*Sargent RL, Craig FE, Swerdlow SH: Comparison of Bcl-2, CD38 and ZAP-70 Expression in Chronic Lymphocytic Leukemia. Int J Clin Exp Pathol. 2009 Jun 162(6):574-82., URL: https://www.ncbi.nlm.nih.gov/pubmed/19636404,*Gachard N, Salviat A, Boutet C, Arnoulet C, Durrieu F, Lenormand B, Leprêtre S, Olschwang S, Jardin F, Lafage-Pochitaloff M, Penther D, Sainty D, Reminieras L, Feuillard J, Béné MC: Multicenter study of ZAP-70 expression in patients with B-cell chronic lymphocytic leukemia using an optimized flow cytometry method. Haematologica. 2008 Feb93(2):215-23., URL: https://www.ncbi.nlm.nih.gov/pubmed/18223290,*Preobrazhensky SN, Bahler DW: Optimization of flow cytometric measurement of ZAP-70 in chronic lymphocytic leukemia. Cytometry B Clin Cytom. 2008 Mar74(2):118-27., URL: https://www.ncbi.nlm.nih.gov/pubmed/17948236,*Del Principe MI, Del Poeta G, Buccisano F, Maurillo L, Venditti A, Zucchetto A, Marini R, Niscola P, Consalvo MA, Mazzone C, Ottaviani L, Panetta P, Bruno A, Bomben R, Suppo G, Degan M, Gattei V, de Fabritiis P, Cantonetti M, Lo Coco F, Del Principe D, Amadori S: Clinical significance of ZAP-70 protein expression in B-cell chronic lymphocytic leukemia. Blood. 2006 Aug 1108(3):853-61., URL: https://www.ncbi.nlm.nih.gov/pubmed/16601244,*Letestu R, Rawstron A, Ghia P, Villamor N, Boeckx N, Boettcher S, Buhl AM, Duerig J, Ibbotson R, Kroeber A, Langerak A, Le Garff-Tavernier M, Mockridge I, Morilla A, Padmore R, Rassenti L, Ritgen M, Shehata M, Smolewski P, Staib P, Ticchioni M, Walker C, Ajchenbaum-Cymbalista F: Evaluation of ZAP-70 expression by flow cytometry in chronic lymphocytic leukemia: A multicentric international harmonization process. Cytometry B Clin Cytom. 2006 Jul 1570(4):309-14., URL: https://www.ncbi.nlm.nih.gov/pubmed/16906588,
• Storage Conditions: Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.