Product Information

Contents: Phycoerythrin (PE) anti-mouse/human ZAP-70 (ZAP70)
Catalog Number: 12-6695
Sizes: 100 tests, 25 ug, 100 ug
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

Staining of permeabilized C57Bl/6 thymocytes (left) and permeabilized Jurkat (right) cells with 0.5 μg of PE Mouse IgG1, K Iso Cntrl (cat. 12-4719) (open histogram) or 0.5 μg of PE 1E7.2 (colored histogram). Total cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-6695	Affinity Purified anti-mouse/human ZAP-70 (ZAP70)	N/A	N/A	IC Flow IP WB
13-6695	Biotin anti-mouse/human ZAP-70 (ZAP70)	N/A	N/A	WB
11-6695	FITC anti-mouse/human ZAP-70 (ZAP70)	488	518	IC Flow
12-6695	PE anti-mouse/human ZAP-70 (ZAP70)	488	575	IC Flow
35-6695	Phycoerythrin-Cy5.5 (PE-Cy5.5) anti-mouse/human ZAP-70 (ZAP70)	488	690	IC Flow
45-6695	PerCP-Cy5.5 anti-mouse/human ZAP-70 (ZAP70)	488	690	IC Flow

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

This 1E7.2 antibody is offered in 2 formats:

- μg size: 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.

- test size: has been pre-titrated and tested by intracellular flow cytometric analysis of mouse thymocytes and human Jurkat cells. This can be used at 20 μl (0.5 μg) per million cells in a 100 μl total staining volume. The pre-titrated test size contains BSA and/or gelatin for protein stabilization.

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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