| Clone: | 1E7.2 |
|---|---|
| Isotype: | Mouse IgG1 |
| Specificity: | The mouse monoclonal antibody 1E7.2 recognizes 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. This product is for research and in vitro experimental use only. It is not to be used for any other commercial purpose. Use of this product to produce products for sale or for therapeutic or drug discovery purposes is prohibited. In order to obtain a license to use this product for commercial purposes, contact The Regents of the Univessity of California. |
| Immunogen: | A KLH-coupled peptide corresponding to amino acids 282-307 of human ZAP-70 |
| Species Reactivity: | Human, Mouse |
| Preparation: | The purified antibody is conjugated with R-Phycoerythrin (PE) under optimum conditions. The conjugate is purified by size-exclusion chromatography and adjusted for direct use. No reconstitution is necessary. |
| Storage Buffer: | The reagent is provided in phosphate buffered saline (PBS) containing 15 mM sodium azide and 0.2% (w/v) high-grade protease free Bovine Serum Albumin (BSA) as a stabilizing agent. |
| Storage / Stability: | Store in the dark at 2-8°C. Do not freeze. Avoid prolonged exposure to light. Do not use after expiration date stamped on vial label. Short-term exposure to room temperature should not affect the quality of the reagent. However, if reagent is stored under any conditions other than those specified, the conditions must be verified by the user. |
| Usage: | The reagent is designed for Flow Cytometry analysis of human blood cells using 20 μl reagent / 100 μl of Permeabilization Medium (originally 50μl of whole blood; see details on the product page) The content of a vial (0.5 ml) is sufficient for 25 tests. |
| Expiration: | See vial label |
| Lot Number: | See vial label |
| 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. |
| References: |
*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 15;70(4):309-14.
*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 1;108(3):853-61.
*Preobrazhensky SN, Bahler DW: Optimization of flow cytometric measurement of ZAP-70 in chronic lymphocytic
leukemia. Cytometry B Clin Cytom. 2008 Mar;74(2):118-27.
*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 Feb;93(2):215-23.
*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 16;2(6):574-82.
*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 Jun;84(6):513-7.
*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 8;8:23.
*And many other.
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