Product Pathways - Jak/Stat Pathway
Stat3 Antibody #9132
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IHC-P ChIP | H M R (B) | Endogenous | 79, 86 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
R=Rat
B=Bovine
Species cross-reactivity is determined by Western blot.
Protocols
Specificity / Sensitivity
Stat3 Antibody detects endogenous levels of total Stat3 protein.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of mouse Stat3. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or IFN-alpha treated (100 ng/ml for 5 min.), using Stat3 Antibody (left) or Phospho-Stat3 (Tyr705) Antibody #9131(right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear and cytoplasmic localization, using Stat3 Antibody.
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HepG2 cells starved overnight and treated with IL-6 (100ng/ml) for 30 minutes, and either 10 μl of Stat3 Antibody #9132 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 primers specific for the c-FOS gene (left) and the c-MYC (right) gene, and the heterochromatic α Satellite repeat element (both). 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. (1999) J. Recept. Signal Transduct. Res. 19, 75-120.
- Takeda, K. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 3801-3804.
- Catlett-Falcone, R. et al. (1999) Immunity 10, 105-115.
- Garcia, R. and Jove, R. (1998) J. Biomed. Sci. 5, 79-85.
- Bromberg, J.F. et al. (1999) Cell 98, 295-303.
- Darnell Jr., J.E. et al. (1994) Science 264, 1415-1421.
- Ihle, J.N. (1995) Nature 377, 591-594.
- Wen, Z. et al. (1995) Cell 82, 241-250.
- Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50.
- Biethahn, S. et al. (1999) Exp. Hematol. 27, 885-894.
Application References
- Bromberg, J. F. et al. (1999) Stat3 as an oncogene. Cell 98, 295-303. Applications: Western Blotting
- Craig, R. et al. (2000) p38 MAPK and NF-kappa B collaborate to induce interleukin-6 gene expression and release. J. Biol. Chem. 275, 23814-23824. Applications: IP 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
- 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
- Ram, P. T. et al. (2000) Stat3-mediated transformation of NIH-3T3 cells by the constitutively active Q205L Galphao protein. Science 287, 142-144. Applications: Western Blotting
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Companion Products
- 9134 Phospho-Stat3 (Ser727) Antibody
- 9914 Phospho-Stat Antibody Sampler Kit
- 9138 Phospho-Stat3 (Tyr705) (3E2) Mouse mAb
- 4904 Stat3 (79D7) Rabbit mAb
- 9139 Stat3 (124H6) Mouse mAb
- 9939 Stat Antibody Sampler Kit
- 9145 Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb
- 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
- 8105 SignalSlide® Phospho-Stat1/3/5 IHC Controls
- 8112 SignalStain® Antibody Diluent
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.