Phospho-Stat3 (Tyr705) Antibod

9131:

ChIP Agarose , ChIP Magnetic , Immunofluorescence* , Immunoprecipitation , Western Blotting

* Product-specific protocol.

Phospho-Stat3 (Tyr705) Antibody detects endogenous levels of Stat3 only when phosphorylated at Tyr705. The antibody does not cross-react with other Stat proteins when phosphorylated on the corresponding tyrosine residue, but has been shown to cross-react with Phospho-EGFR.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr705 of mouse Stat3. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from CNTF-treated SK-N-MC cells and EGF-treated A431 cells, using Phospho-Stat3 (Tyr705) Antibody (upper) or Stat3 Antibody #9132 (lower).

IP

Immunoprecipitation of extracts from CNTF-treated 35S-labeled SK-N-MC cells, using Phospho-Stat3 (Tyr705) Antibody (upper) or Stat3 Antibody #9132 (lower).

IF-IC

Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or IFNalpha-treated (right), using Phospho-Stat3 (Tyr705) Antibody (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red).

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and either 10 μl of Phospho-Stat3 (Tyr705) 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 c-Fos Promoter Primers #4663, 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 Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
7. Ihle, J.N. (1995) Nature 377, 591-4.
8. Wen, Z. et al. (1995) Cell 82, 241-50.
9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

• Bartoli, M. et al. (2000) Vascular endothelial growth factor activates STAT proteins in aortic endothelial cells. J. Biol. Chem. 275, 33189-33192. Applications: IC-IF Western Blotting
• Campbell, C. L. et al. (2001) Increased expression of the interleukin-11 receptor and evidence of STAT3 activation of prostate carcinoma. Am. J. Path. 158, 25-32. Applications: Western Blotting
• Mitsui, K. et al. (2003) The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631-642. Applications: Western Blotting
• Guo, Z. et al. (2004) CDC91L1 (PIG-U) is a newly discovered oncogene in human bladder cancer. Nat. Med. 10, 374-381. Applications: IC-IF Western Blotting
• Butzkueven, H. et al. (2002) LIF receptor signaling limits immune-mediated demyelination by enhancing oligodendrocyte survival. Nat. Med. 8, 613-619. Applications: Western Blotting
• Bild, A. H. et al. (2002) Cytoplasmic transport of Stat3 by receptor-mediated endocytosis. EMBO J. 21, 3255-3263. Applications: Western Blotting
• Ducy, P. et al. (2000) Leptin inhibits bone formation through a hypothalamic relay: a central control of bone mass. Cell 100, 197-207. Applications: Western Blotting
• Fujimoto, M. et al. (2000) Defective thymocyte development and perturbed homeostasis of T cells in STAT-induced STAT inhibitor-1/suppressors of cytokine signaling-1 transgenic mice. J. Immunol. 165, 1799-1806. Applications: Western Blotting
• Hideshima, T. et al. (2001) The proteasome inhibitor PS-341 inhibits growth, induces apoptosis, and overcomes drug resistance in human multiple myeloma cells. Cancer Res. 61, 3071-3076. Applications: Western Blotting
• Stairs, D.B. et al. (2011) Cancer Cell 19, 470-83. Applications: Western Blotting
• Zhang, W.N. et al. (2012) J Biol Chem 287, 382-92. 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 !

• 9130 PhosphoPlus® Stat3 (Tyr705) Antibody Kit
• 9132 Stat3 Antibody
• 9133 Stat3 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
• 9138 Phospho-Stat3 (Tyr705) (3E2) Mouse mAb
• 9145 Phospho-Stat3 (Tyr705) (D3A7) XP^® Rabbit mAb

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