Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Stat3 (124H6) Mouse mAb #9139

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC F ChIP H M R Mk Endogenous 79, 86 Mouse IgG2a

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Stat3 (124H6) Mouse mAb detects endogenous levels of total Stat3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of mouse Stat3.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, NIH/3T3, PC12 and COS cells, using Stat3 (124H6) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma (left), showing nuclear and cytoplasmic staining, and human lung carcinoma (right), showing cytoplasmic staining, using Stat3 (124H6) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Hela cells, untreated (blue) or IFN-alpha-treated (green), using Stat3 (124H6) Mouse mAb compared with a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or IFNalpha-treated (right) and labeled with Stat3 (124H6) Mouse mAb (green).

Chromatin IP

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 (124H6) Mouse mAb #9139 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), the c-MYC gene (right), 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).

  1. Heim, M.H. (1999) J. Recept. Signal Transduct. Res. 19, 75-120.
  2. Takeda, K. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 3801-3804.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-115.
  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 Jr., J.E. et al. (1994) Science 264, 1415-1421.
  7. Ihle, J.N. (1995) Nature 377, 591-594.
  8. Wen, Z. et al. (1995) Cell 82, 241-250.
  9. Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp. Hematol. 27, 885-894.

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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