Product Pathways - Jak/Stat Pathway
Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145
|W IP IHC-P IHC-F IF-IC F ChIP||H M R Mk (Hm) (B) (Pg) (Hr)||Endogenous||79, 86||Rabbit IgG|
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey B=Bovine 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.
* Product-specific protocol.
Specificity / Sensitivity
Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb detects endogenous levels of Stat3 only when phosphorylated at tyrosine 705. This antibody does not cross-react with phospho-EGFR or the corresponding phospho-tyrosines of other Stat proteins.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr705 of mouse Stat3.
Western blot analysis of extracts from IFN-alpha treated Jurkat cells and HeLa cells (left), as well as EGF treated A431 cells (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb. Note that the basal phospho-Stat3 in A431 is detected by the antibody.
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine, using Phosho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin embedded human breast carcinoma, specifically endothelial cells, untreated (left) or lambda phosphatase treated (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
Immunnohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
Immunohistochemical analysis using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb on SignalSlide® Phospho-Stat1/3/5 IHC Controls #8105 (paraffin-embedded HeLa cell pellets, untreated (left), treated with Human Interferon-α1 (hIFN-α1) #8927 (middle), or treated with Human Epidermal Growth Factor (hEGF) #8916 (right).
Immunohistochemical analysis of frozen H1650 xenograft using Phospho-Stat3 (Tyr705)(D3A7) XP® Rabbit mAb.
Flow cytometric analysis of HeLa cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells, IFN-alpha treated (left) or untreated (right), labeled with Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (green).
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 10 μl of Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb 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). 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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- 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.
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- 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.
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- Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
- Tripathi, R.B. and McTigue, D.M. (2008) J Comp Neurol 510, 129-44. Applications: IC-IF
- Ernst, M.B. et al. (2009) J Neurosci 29, 11582-93. Applications: IHC-F (floating)
- Bauer, S. and Patterson, P.H. (2006) J Neurosci 26, 12089-99. Applications: IHC-FL (floating/frozen)
- Yao, Z. et al. (2010) Proc Natl Acad Sci U S A 107, 15535-40. Applications: Western Blotting
- Surdziel, E. et al. (2011) Blood 117, 4338-48. Applications: Western Blotting
- Stairs, D.B. et al. (2011) Cancer Cell 19, 470-83. Applications: IC-IF
- Anand, S. et al. (2011) Blood 118, 1610-21. Applications: Western Blotting
- Mainardi, M. et al. (2010) Proc Natl Acad Sci U S A 107, 16673-8. Applications: IF-F
- Matsuyama, H. et al. (2011) Blood 118, 6881-92. Applications: Western Blotting
- Ahluwalia, M. et al. (2010) J Thromb Haemost 8, 2252-61. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.