Stat1 Antibody

9172:

ChIP Agarose , ChIP Magnetic , Immunoprecipitation , Western Blotting

Stat1 Antibody detects endogenous levels of total Stat1 protein. The antibody detects both Stat1alpha (91kDa) and Stat1beta (84 kDa) isoforms.

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

Western Blotting

Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with non-targeted (control) siRNA or transfection with Stat1 siRNA. Stat1 was detected using Stat1 Antibody #9172, and p42 was detected using p42 MAPK Antibody #9108. The Stat1 Antibody confirms silencing of Stat1 expression, and the p42 MAPK Antibody is used to control for protein loading and siRNA specificity.

Western Blotting

Western blot analysis of extracts from SK-MEL-28 cells, untreated or IFN-alpha-treated (100 ng/ml), using Phospho-Stat1 (Tyr701) Antibody #9171 (upper) or Stat1 Antibody (lower).

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 10 μl of Stat1 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.

• Chen, G. et al. (2001) Expression of the transcription factor STAT-1 alpha in insulinoma cells protects against cytotoxic effects of multiple cytokines. J. Biol. Chem. 276, 766-772. Applications: Western Blotting
• Hutt, J. A. et al. (2000) Signal transducer and activator of transcription 3 activates CCAAT enhancer-binding protein delta gene transcription in G0 growth-arrested mouse mammary epithelial cells and in involuting mouse mammary gland. J. Biol. Chem. 275, 29123-29131. Applications: IP Western Blotting
• Mahboubi, K. and Pober, J.S. (2002) Activation of signal transducer and activator of transcription 1 (STAT1) is not sufficient for the induction of STAT1-dependent genes in endothelial cells. J. Biol. Chem. 277, 8012-8021. Applications: Western Blotting
• Peterson, W. M. et al. (2000) Ciliary neurotrophic factor and stress stimuli activate the Jak-STAT pathway in retinal neurons and glia. J. Neurosci. 20, 4081-4090. Applications: Western Blotting
• Suzuki, K. et al. (1999) Activation of target-tissue immune-recognition molecules by double-stranded polynucleotides. Proc. Natl. Acad. Sci. USA 96, 2285-2290. Applications: Western Blotting
• Zimnik, S. et al. (2009) Nucleic Acids Res 37, e30. Applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

• 9171 Phospho-Stat1 (Tyr701) Antibody
• 9173 Stat1 Control Cell Extracts
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9170 PhosphoPlus® Stat1 (Tyr701) Antibody Kit
• 9914 Phospho-Stat Antibody Sampler Kit
• 9175 Stat1 (42H3) Rabbit mAb
• 9939 Stat Antibody Sampler Kit

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