Phospho-Stat1 (Tyr701) Antibod

9171:

ChIP Agarose , ChIP Magnetic , Immunoprecipitation , Western Blotting

Phospho-Stat1 (Tyr701) Antibody detects endogenous levels of Stat1 only when phosphorylated at tyrosine 701. The antibody detects phosphorylated tyrosine 701 of p91 Stat1 and also the p84 splice variant. It does not cross-react with the corresponding phospho-tyrosines of other Stat proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr701 of human Stat1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or IFN-alpha-treated (100 ng/ml), using Phospho-Stat1 (Tyr701) Antibody.

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 20 μl of Phospho-Stat1 (Tyr701) Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP^® Human TAP1 Promoter Primers #5148, and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.

1. Heim, M.H. (1999) J. Recept. Signal. Transduct. Res. 19, 75-120.
2. Durbin, J.E. et al. (1996) Cell 84, 443-450.
3. Meraz, M.A. et al. (1996) Cell 84, 431-442.
4. Ihle, J.N. et al. (1994) Trends Biochem. Sci. 19, 222-227.
5. Frank, D.A. (1999) Mol. Med. 5, 432-456.
6. Wen, Z. et al. (1995) Cell 82, 241-250.

• Begitt, A. et al. (2000) Nucleocytoplasmic translocation of Stat1 is regulated by a leucine-rich export signal in the coiled-coil domain. Proc. Natl. Acad. Sci. USA 97, 10418-10423. Applications: Western Blotting
• Gamero, A.M. and Larner, A.C. (2000) Signaling via the T cell antigen receptor induces phosphorylation of Stat1 on serine 727. J. Biol. Chem. 275, 16574-16578. Applications: Western Blotting
• McBride, K. M. et al. (2000) Nuclear export signal located within the DNA-binding domain of the STAT1 transcription factor. EMBO J. 19, 6196-6206. Applications: Western Blotting
• Spagnoli, A. et al. (2002) Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. J. Biol. Chem. 277, 18860-18867. Applications: IC-ABC Western Blotting
• Hirata, A. et al. (2002) ZD1839 (Iressa) induces antiangiogenic effects through inhibition of epidermal growth factor receptor tyrosine kinase. Cancer Res. 62, 2554-2560. Applications: Western Blotting
• Hayashi, T. et al. (2002) Regulation of receptor activator of NF-kappaB ligand-induced osteoclastogenesis by endogenous interferon-beta (INF-beta) and suppressors of cytokine signaling (SOCS). J. Biol. Chem. 277, 27880-27886. Applications: Western Blotting
• Mowen, K. A. et al. (2001) Arginine methylation of STAT1 modulates IFN alpha/beta-induced transcription. Cell 104, 731-741. Applications: Western Blotting
• Teague, T. K. et al. (2000) Activation-induced inhibition of interleukin 6-mediated T cell survival and signal transducer and activator of transcription 1 signaling. J. Exp. Med. 191, 915-926. Applications: Western Blotting
• Lee, J. et al. (2011) Oncogene , . Applications: Western Blotting
• Zimnik, S. et al. (2009) Nucleic Acids Res 37, e30. Applications: Western Blotting

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For Research Use Only. Not For Use In Diagnostic Procedures.