antibody

ZAP 70 (Tyr319) Syk (Tyr352), phosphorylated (TCR Zeta Associated Protein kinase 70kD)

Z0300-06

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

ZAP 70 (Tyr319) Syk (Tyr352), phosphorylated (TCR Zeta Associated Protein kinase 70kD)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

ZAP70, a Syk family protein tyrosine kinase expressed in T and NK cells, plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement. Following TCR engagement, ZAP70 is rapidly phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a transphosphorylation by the Src family tyrosine kinase Lck. Tyrosine phosphorylation of ZAP70 correlates with its increased kinase activity and couples to downstream signaling events. Phosphorylation of Tyr319 is required for the assembly of a ZAP70-containing signaling complex that leads to the activation of the PLC-gamma1-dependent and Ras-dependent signaling cascades in antigen-stimulated T cells. The orthologous Tyr352 residue in Syk is also involved in the association of PLC-gamma1. Phosphorylation of ZAP70 is crucial to the recruitment of additional molecules, such as Vav, Ras, and PLC gamma-1, to the membrane-associated signaling complex and for the induction of downstream signaling events. Mutation of the ZAP70 genes results in a form of Severe Combined Immunodeficiency Syndrome (SCID) in humans. Recent evidence has suggested that ZAP70 should be considered as an effective biomarker in Chronic Lymphocytic Leukemia (CLL). Applications: Suitable for use in Western Blot, Immunocytochemistry and Flow Cytometry. Other applications have not been tested. Recommended Dilution: Western Blotting: 1:1000 Immunoprecipitation: 1:100 Immunocytochemistry (for IF): 1:100 Flow Cytometry: 1:25 Optimal dilutions to be determined by the researcher. Molecular Weight: 70kD (Zap-70), 72kD (Syk) Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Hu

Synthetic phospho-peptide corresponding to residues surrounding Tyr319 of human Zap-70. Species sequence homology: mouse, rat, hamster, monkey, bovine, canine, porcine and equine

Detects endogenous levels of human Zap-70 only when phosphorylated at Tyr319. Crossreacts with endogenous levels of Syk when phosphorylated at Tyr352.

Purified by Protein A and peptide affinity chromatography.

Supplied as liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.

Not determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not determined Form: Supplied as liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Tyr319 of human Zap-70. Species sequence homology: mouse, rat, hamster, monkey, bovine, canine, porcine and equine Specificity: Detects endogenous levels of human Zap-70 only when phosphorylated at Tyr319. Crossreacts with endogenous levels of Syk when phosphorylated at Tyr352. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

FC IC IF WB

-20°C

1. Chu, D. H. et al. (1998) Immunol. Rev. 165, 167–180. 2. Iwashima, M. et al. (1994) Science 263, 1136–1139. 3. Neumeister, E. N. et al. (1995) Mol. Cell. Biol. 15, 3171–3178. 4. Chan, A. C. et al. (1995) EMBO J. 14, 2499–2508. 5. Williams, B. L. et al. (1999) EMBO J. 18, 1832–1844. 6. Di Bartolo, V. et al. (1999) J. Biol. Chem. 274, 6285–6294. 7. Law, C. L. et al. (1996) Mol. Cell. Biol. 16, 1305–1315. 8. Guntermann, C. and Alexander, D.R. (2002) J. Immunol. 168, 4420–4429.

https://usbio-images.r.worldssl.net/prodimages/9/Z0300-06_1.jpg

Flow cytometric analysis of Jurkat cells, untreated (blue) or CD3 treated (green), using Phospho-Zap70 (Tyr319)/Syk (Tyr352) Antibody compared to a nonspecific negative control antibody (red).