Anti-ZAP70 Purified

Anti-ZAP70 Purified
Regulatory status
RUO
Antigen
ZAP70
Clone
1E7.2
Format
Purified
Reactivity
Human, Mouse
Variant
0.1 mg
11-665-C100
Ask for delivery term HERE
150.00 USD

0.025 mg
11-665-C025
Delivery 1 week
75.00 USD
Variant
0.1 mg
11-261-C100
In stock
150.00 USD

0.1 mg
11-261-C100
In stock
75.00 USD
Product details
Description
Images
References
Isotype
Mouse IgG1 kappa
Specificity
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".
Application details
Flow cytometry: Recommended dilution for purified antibody: 1 μg/ml, intracellular staining. Recommended protocol for primary antibody conjugates using ADG Fix and Perm kit: 1) Add 50 μl peripheral blood to a 5 ml FACS tube; 2) Proceed surface staining with appropriate amount of surface antibody (20 min at room temperature); 3) Add 100 μl of Reagent A (Fixation Medium, stored and used at room temperature); 4) Incubate for 15 minutes at room temperature; 5) Add 4 ml PBS buffer and centrifuge at 300 g for 5 minutes at room temperature; 6) Remove supernatant and add to cells pellet 100 μl Reagent B (Permeabilization Medium) and 20 μl of ZAP-70-PE antibody or 10 μl of ZAP-70-FITC antibody (red blood cells are not lysed yet); 7) Vortex at low speed for 1-2 seconds (pellet must be resuspended); 8) Incubate for 15 minutes at room temperature; 9) Add to cells mixture of 1 ml Reagent B and 3 ml PBS buffer and centrifuge at 300 g for 5 minutes at room temperature; 10) Remove supernatant and resuspend cells in 100 μl PBS buffer; 11) Analyze immediately.
Western blotting: Recommended dilution: 1-2 μg/ml, positive control: MOLT-4 cells.
Reactivity
Human, Mouse
Immunogen
A KLH-coupled peptide corresponding to amino acids 282-307 of human ZAP70
Concentration
1 mg/ml
Preparation
Purified by protein-A affinity chromatography.
Formulation
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Storage and handling
Store at 2-8°C. Do not freeze.
Exbio licence note
The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.
Licence notice
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 University of California.
Other names
ZAP-70, ADMIO2
Antigen description
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.
Entrez Gene ID 22637
UniProt ID P43404
11-665_WB
Western blotting analysis of human ZAP70 using mouse monoclonal antibody 1E7.2 on lysates of MOLT-4 cell line and HEK293T/17 cell line (ZAP70 non-expressing cell line; negative control) under non-reducing and reducing conditions. Nitrocellulose membrane was probed with 2 µg/ml of mouse anti-ZAP70 monoclonal antibody 1E7.2 followed by IRDye800-conjugated anti-mouse IgG1 secondary antibody. A specific band was detected for ZAP70 protein at approximately 70 kDa.

Product specific references:

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.
PubMed
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.
PubMed
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.
PubMed
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.
PubMed
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.
PubMed
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.
PubMed
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.
PubMed
Variant
0.1 mg
11-665-C100
Ask for delivery term HERE
150.00 USD

0.025 mg
11-665-C025
Delivery 1 week
75.00 USD
Variant
0.1 mg
11-261-C100
In stock
150.00 USD

0.1 mg
11-261-C100
In stock
75.00 USD

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