| Application |
Dilution |
Species-Reactivity |
Sensitivity |
Isotype |
| W |
|
Human, Mouse, Rat, Hamster, Monkey |
Endogenous |
Rabbit IgG |
| IP |
|
| IHC-P |
|
| IF-IC |
|
| ChIP |
|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Homology
Species predicted to react based on 100% sequence homology:
Pig.
Directions For Use
For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP.
This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Specificity / Sensitivity
Stat3α (D1A5) XP® Rabbit mAb recognizes endogenous levels of total Stat3α protein. This antibody does not cross-react with Stat3β.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Stat3α protein.
Western Blotting
Western blot analysis of extracts from various cell lines using Stat3α (D1A5) XP® Rabbit mAb (top), Stat3 (124H6) Mouse mAb #9139 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded cell pellets, HeLa (left) or PC-3 (right), using Stat3α (D1A5) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Stat3α (D1A5) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/mL, 30 min; right) using Stat3α (D1A5) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml, 30 min), and either Stat3α (D1A5) XP® Rabbit mAb or 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.
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). 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.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoSitePlus® protein, site, and accession data:
