Species predicted to react based on 100% sequence homology: Chicken, Bovine, Dog.

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1 1 of 5 11/1/ :25 PM Product Pathways - Jak/Stat Pathway Phospho-Stat3 (Tyr705) Antibody #9131 Have you tried your application using our XP monoclonal antibodies? Try products: 9145 PhosphoSitePlus protein, site, and accession data: STAT3 No. Size Price 9131L 300 ul ( 30 western blots ) please select country 9131S 100 ul ( 10 western blots ) please select country 9131 carrier free & custom formulation / quantity request Secondary Antibodies Selection Write a Review Already purchased this product? Write a Review. Application Dilution Species-Reactivity Sensitivity MW (kda) Source W 1:1000 Human, Mouse, Rat, Monkey Endogenous 79, 86 Rabbit IP 1:100 ChIP 1:50 Species cross-reactivity is determined by western blot. Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP Homology Species predicted to react based on 100% sequence homology: Chicken, Bovine, Dog. Protocols 9131: Western Blotting, Immunoprecipitation, Immunofluorescence*, ChIP Magnetic, ChIP Agarose * Product-specific protocol. Specificity / Sensitivity 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

2 2 of 5 11/1/ :25 PM corresponding tyrosine residue, but has been shown to cross-react with Phospho-EGFR. Source / Purification 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 IP 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). 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

3 Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 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. Background 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). Research studies have shown that 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) Heim, M.H. (2001) J Recept Signal Transduct Res 19, Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, of 5 11/1/ :25 PM

4 Catlett-Falcone, R. et al. (1999) Immunity 10, Garcia, R. and Jove, R. (1998) J Biomed Sci 5, Bromberg, J.F. et al. (1999) Cell 98, Darnell, J.E. et al. (1994) Science 264, Ihle, J.N. (1995) Nature 377, Wen, Z. et al. (1995) Cell 82, Yokogami, K. et al. (2000) Curr Biol 10, Biethahn, S. et al. (1999) Exp Hematol 27, Application References Bartoli, M. et al. (2000). J. Biol. Chem. 275, Applications: Western Blotting, IC-IF. Campbell, C. L. et al. (2001). Am. J. Path. 158, Applications: Western Blotting. Mitsui, K. et al. (2003) Cell 113, Applications: Western Blotting. Guo, Z. et al. (2004) Nat. Med. 10, Applications: Western Blotting, IC-IF. Butzkueven, H. et al. (2002). Nat. Med. 8, Applications: Western Blotting. Bild, A. H. et al. (2002). EMBO J. 21, Applications: Western Blotting. Ducy, P. et al. (2000). Cell 100, Applications: Western Blotting. Fujimoto, M. et al. (2000). J. Immunol. 165, Applications: Western Blotting. Hideshima, T. et al. (2001). Cancer Res. 61, Applications: Western Blotting. Stairs, D.B. et al. (2011) Cancer Cell 19, Applications: Western Blotting. Marotta, L.L. et al. (2011) J Clin Invest 121, Applications: Western Blotting, IF-IC (In Cells). Zhang, W.N. et al. (2012) J Biol Chem 287, Applications: Western Blotting. Fiedler, J. et al. (2012) Haematologica 97, 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! Companion Products 9130 PhosphoPlus Stat3 (Tyr705) Antibody Kit 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 X LumiGLO Reagent and 20X Peroxide 9138 Phospho-Stat3 (Tyr705) (3E2) Mouse mab 4 of 5 11/1/ :25 PM

5 5 of 5 11/1/ :25 PM 9145 Phospho-Stat3 (Tyr705) (D3A7) XP Rabbit mab For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc Cell Signaling Technology, Inc. contact privacy policy browsers & accessibility sitemap careers

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