Product Pathways - Jak/Stat Pathway
Stat3 (D3Z2G) Rabbit mAb #12640
|12640S||100 µl (10 western blots)||---||In Stock||---|
|12640||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||79, 86||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Bovine, Pig.
* Product-specific protocol.
Specificity / Sensitivity
Stat3 (D3Z2G) Rabbit mAb recognizes endogenous levels of total Stat3 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly700 of human Stat3 protein.
Western blot analysis of extracts from various cell lines using Stat3 (D3Z2G) Rabbit mAb (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower). PC-3 cells are negative for Stat3.
Immunoprecipitation of Stat3 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Stat3 (D3Z2G) Rabbit mAb (lane 3). Lane 1 is 10% input.
Flow cytometric analysis of of PC-3 (blue) and HeLa (green) cells using Stat3 (D3Z2G) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of serum-starved HeLa cells, untreated (upper left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; upper right), and serum-starved PC-3 cells, untreated (lower right) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; lower right), using Stat3 (D3Z2G) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml) for 30 minutes, and either 10 μl of Stat3 (D3Z2G) 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). Research studies have shown that 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. (2001) J Recept Signal Transduct Res 19, 75-120.
- 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.
- Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
- 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.
- Wen, Z. et al. (1995) Cell 82, 241-50.
- Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
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For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
SimpleChIP® is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.