Product Pathways - Jak/Stat Pathway

Stat3 Antibody #9132

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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

9132:: ChIP Agarose, ChIP Magnetic, IHC / Paraffin, Immunoprecipitation, Western Blotting

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).

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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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.