Product Pathways - Jak/Stat Pathway
Phospho-Stat3 (Ser727) Antibody #9134
|9134L||300 µl (30 western blots)||---||In Stock||---|
|9134S||100 µl (10 western blots)||---||In Stock||---|
|9134P||40 µl (4 western blots)||---||In Stock||---|
|9134||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat||Endogenous||86||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Hamster, Monkey, Bovine, Dog, Pig, Horse.
* Product-specific protocol.
Specificity / Sensitivity
Phospho-Stat3 (Ser727) Antibody recognizes endogenous levels of Stat3 only when phosphorylated at Ser727. The antibody does not significantly cross-react with the corresponding phospho-serines of other Stat proteins.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser727 of mouse Stat3. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from A172 cells, untreated (-) or UV-treated (100 mJ, 30 min; +) with or without λ phosphatase (+), using Phospho-Stat3 (Ser727) Antibody (upper) and Stat3 (79D7) Rabbit mAb #4904 (lower).
Confocal immunofluorescent analysis of HeLa cells, untreated (left), IL-6 treated (center) or IL-6 and phosphatase treated (right), using Phospho-Stat3 (Ser727) Antibody (green) and Pan-Keratin (C11) Mouse mAb #4545 (red).
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, 30 min), and either 20 μl of Phospho-Stat3 (Ser727) 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). 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).
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- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
- Bromberg, J. F. et al. (1999) Cell 98, 295-303. Applications: Western Blotting.
- Wu, Y.Y. and Bradshaw, R.A. (2000) J. Biol. Chem. 275, 2147-2156. Applications: Western Blotting.
- Teague, T. K. et al. (2000) J. Exp. Med. 191, 915-926. Applications: Western Blotting.
- Matsuda, T. et al. (1999) EMBO J. 18, 4261-4269. Applications: Western Blotting.
- Schuringa, J. J. et al. (2001) . Oncogene 20 (38), 5350-5358. Applications: Western Blotting.
- Simon, A. R. et al. (2000) Science 290, 144-147. Applications: Western Blotting.
- Lee, J. et al. (2011) Oncogene , . Applications: Western Blotting.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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SimpleChIP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.