Contents: Fluorescein isothiocyanate (FITC) anti-mouse/human ZAP-70 (ZAP70)
Catalog Number: 11-6695
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN3
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: 1E7.2
Isotype: Mouse IgG1, κ
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Staining of permeabilized Jurkat (left) and permeabilized C57Bl/6 thymus (right) cells with 0.5 μg of FITC Mouse IgG1, K Iso Cntrl (I/C Staining) (cat. 11-4718) (open histogram) or 0.5 μg of FITC 1E7.2 (colored histogram). Total cells were used for analysis.
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Questions? Please consult our answers to frequently asked questions at
http://www.ebioscience.com/faq.
Description
The 1E7.2 antibody reacts with human and mouse ZAP-70, the TCRζ-associated protein-70. ZAP-70 is a cytosolic protein tyrosine kinase (PTK) and a member of the Syk family of proteins. It is expressed in T and NK cells and is required for TCR signaling and development. ZAP-70 interacts with the TCR complex by binding to tyrosine-phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMs) present in the invariant subunits of the TCR complex. Following activation, ZAP-70 is phosphorylated on several tyrosine residues by two mechanisms; an autophosphorylation and a transphosphorylation by the Src family tyrosine kinase Lck1-3. Tyrosine phosphorylation of ZAP-70 correlates to its increased kinase activity and triggers downstream signaling events. Mutations in ZAP-70 have been shown to result in a form of Severe Combined Immunodeficiency Syndrome (SCID) in humans. 1E7.2 was generated against a KLH-peptide sequence corresponding to the human ZAP-70 amino acid residues 282-307. While ZAP-70 is normally expressed in T and NK cells, several recent studies have also shown high correlation of ZAP-70 positive expression with mutated IgVH expression in B-chronic lymphocytic leukemia (CCL). In conclusion, the expression of ZAP-70, which can be measured by intracellular flow cytometry, may serve as a prognostic marker for B-CLL.
Applications Reported
The 1E7.2 antibody has been reported for use in intracellular flow cytometric analysis. This product is for
in vitro research use only. It is not to be used for commercial purposes. Use of this product to produce products for sale or for diagnostic, 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.
Please visit the following website for a general intracellular staining protocol:
http://www.ebioscience.com/ebioscience/appls/FCI.htm
Applications Tested
The 1E7.2 antibody has been tested by intracellular flow cytometric analysis of mouse thymocytes and human Jurkat cells. This can be used at less than or equal to 1 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Qian D, Lev S, van Oers NS, Dikic I, Schlessinger J, Weiss A. 1997. Tyrosine phosphorylation of Pyk2 is selectively regulated by Fyn during TCR signaling. J Exp Med. 185(7):1253-9.
Qian D, Mollenauer MN, Weiss A. 1996. Dominant-negative zeta-associated protein 70 inhibits T cell antigen receptor signaling. J Exp Med. 183(2):611-20.
Orchard JA, Ibbotson RE, Davis Z, Wiestner A, Rosenwald A, Thomas PW, Hamblin TJ, Staudt LM, Oscier DG. 2004. ZAP-70 expression and prognosis in chronic lymphocytic leukaemia. Lancet. Jan 10;363(9403):105-11.
Chen L, Widhopf G, Huynh L, Rassenti L, Rai KR, Weiss A, Kipps TJ. 2002. Expression of ZAP-70 is associated with increased B-cell receptor signaling in chronic lymphocytic leukemia. Blood. Dec 15;100(13):4609-14.
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