Product Pathways - Jak/Stat Pathway
Phospho-Stat3 (Ser727) Antibody #9134
PhosphoSitePlus® protein, site, and accession data: STAT3
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IF-IC ChIP | H M R (Hm) (Mk) (B) (Dg) (Pg) (Hr) | Endogenous | 86 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
R=Rat
Hm=Hamster
Mk=Monkey
B=Bovine
Dg=Dog
Pg=Pig
Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
* Product-specific protocol.
Specificity / Sensitivity
Phospho-Stat3 (Ser727) Antibody detects 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 Blotting
Western blot analysis of extracts from A431 cells, untreated or EGF-treated (100 ng/ml) for the indicated times, using Phospho-Stat3 (Ser727) Antibody (upper) or Stat3 Antibody #9132 (lower).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left), IL-6 treated (center) or IL-6 and phosphatase treated (right), labeled with Phospho-Stat3 (Ser727) Antibody (green) and Pan-Keratin (C11) Mouse mAb #4545 (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 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.
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). 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, 75-120.
- Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
- Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
- Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
- Bromberg, J.F. et al. (1999) Cell 98, 295-303.
- Darnell, J.E. et al. (1994) Science 264, 1415-21.
- Ihle, J.N. (1995) Nature 377, 591-4.
- Wen, Z. et al. (1995) Cell 82, 241-50.
- Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
Application References
- Bromberg, J. F. et al. (1999) Stat3 as an oncogene. Cell 98, 295-303. Applications: Western Blotting
- Wu, Y.Y. and Bradshaw, R.A. (2000) Activation of the Stat3 signaling pathway is required for differentiation by interleukin-6 in PC12-E2 cells. J. Biol. Chem. 275, 2147-2156. 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
- Matsuda, T. et al. (1999) STAT3 activation is sufficient to maintain an undifferentiated state of mouse embryonic stem cells. EMBO J. 18, 4261-4269. Applications: Western Blotting
- Schuringa, J. J. et al. (2001) MEN2A-RET-induced cellular transformation by activation of STAT3. Oncogene 20 (38), 5350-5358. Applications: Western Blotting
- Simon, A. R. et al. (2000) Regulation of STAT3 by direct binding to the rac1 GTPase. Science 290, 144-147. Applications: Western Blotting
- Lee, J. et al. (2011) Oncogene , . Applications: Western Blotting
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Companion Products
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- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 4904 Stat3 (79D7) Rabbit mAb
- 9139 Stat3 (124H6) Mouse mAb
For Research Use Only. Not For Use In Diagnostic Procedures.