Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb #4113

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC F H M R Mk Endogenous 79, 86 Mouse IgG1

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
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 (Tyr705) (M9C6) Mouse mAb detects endogenous levels of Stat3 only when phosphorylated at Tyr705. 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 Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with IFN-α (100 ng/ml) for 5 minutes, using Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb (left) or Stat3 Antibody #9132 (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical anlysis of paraffin-embedded human breast carcinoma using Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 786-O cells (SignalSlide® Phospho-Stat1/3/5 IHC Controls #8105), untreated (left) or IFN-α-treated (right), using Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or IFN-alpha treated (right), using Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

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).

  1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
  2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
  6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  7. Ihle, J.N. (1995) Nature 377, 591-4.
  8. Wen, Z. et al. (1995) Cell 82, 241-50.
  9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

Application References

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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.

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