Product Pathways - Jak/Stat Pathway
Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb #9145
This XP™ monoclonal antibody was developed using our eXceptional Monoclonal Technology (XMT™).
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IHC-F IF-IC F ChIP | H M R Mk | Endogenous | 79, 86 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
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
- 9145:
- ChIP Agarose, ChIP Magnetic, Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
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 Blotting
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.
IHC-P (paraffin)
Immunohistochemical analysis using Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb on SignalSlide (TM) Phospho-Stat 1/3/5 IHC Controls #8105 (paraffin-embedded 786-0 cells, untreated (left) or IFN-a treated (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine, using Phosho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb.
IHC-P (paraffin)
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.
IHC-P (paraffin)
Immunnohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb.
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft using Phospho-Stat3 (Tyr705)(D3A7) XP™ Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of HeLa cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb.
IF-IC
Confocal immunofluorescent analysis of HeLa cells, IFN-alpha treated (left) or untreated (right), labeled with Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb (green).
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 Phospho-Stat3 (Tyr705) (D3A7) XP™ Rabbit mAb #9145 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).
- Heim, M.H. (1999) J. Recept. Signal Transduct. Res. 19, 75-120.
- Takeda, K. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 3801-3804.
- Catlett-Falcone, R. et al. (1999) Immunity 10, 105-115.
- Garcia, R. and Jove, R. (1998) J. Biomed. Sci. 5, 79-85.
- Bromberg, J.F. et al. (1999) Cell 98, 295-303.
- Darnell Jr., J.E. et al. (1994) Science 264, 1415-1421.
- Ihle, J.N. (1995) Nature 377, 591-594.
- Wen, Z. et al. (1995) Cell 82, 241-250.
- Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50.
- Biethahn, S. et al. (1999) Exp. Hematol. 27, 885-894.
Application References
- 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)
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9131 Phospho-Stat3 (Tyr705) Antibody
- 9132 Stat3 Antibody
- 9133 Stat3 Control Cell Extracts
- 9134 Phospho-Stat3 (Ser727) Antibody
- 9136 Phospho-Stat3 (Ser727) (6E4) Mouse mAb
- 9138 Phospho-Stat3 (Tyr705) (3E2) Mouse mAb
- 9139 Stat3 (124H6) Mouse mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 1195 Phospho-Stat3 (Tyr705) Blocking Peptide
- 8105 SignalSlide® Phospho-Stat1/3/5 IHC Controls
- 8112 SignalStain® Antibody Diluent
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.
