Product Pathways - Jak/Stat Pathway
Phospho-Stat5 (Tyr694) Antibody #9351
|9351L||300 µl (30 western blots)||---||In Stock||---|
|9351S||100 µl (10 western blots)||---||In Stock||---|
|9351||carrier free and custom formulation / quantity||email request|
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Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, F=Flow Cytometry, ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Rat, Bovine.
Specificity / Sensitivity
Phospho-Stat5 (Tyr694) Antibody detects endogenous levels of Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699. The antibody does not cross-react with the corresponding phospho-tyrosine residues of other Stat proteins, but does cross-react with EGFR.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr694 of mouse Stat5a. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from TF-1 cells, treated with GM-CSF (25 ng/ml) for the indicated times, using Phospho-Stat5 (Tyr694) Antibody (upper) or Stat5 antibody (lower).
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue), using Phospho-Stat5 (Tyr694) Antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 20 μl of Phospho-Stat5 (Tyr694) Antibody 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 SimpleChIP® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. 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.
Stat5 is activated in response to a wide variety of ligands including IL-2, GM-CSF, growth hormone and prolactin. Phosphorylation at Tyr694 is obligatory for Stat5 activation (1,2). This phosphorylation is mediated by Src upon erythropoietin stimulation (3). Stat5 is constitutively active in some leukemic cell types (4). Phosphorylated Stat5 is found in some endothelial cells treated with IL-3, which suggests its involvement in angiogenesis and cell motility (5). Stat5a and Stat5b are independently regulated and activated in various cell types. For instance, interferon treatment predominantly activates Stat5a in U-937 cells and Stat5b in HeLa cells (6).
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- Wakao, H. et al. (1994) EMBO J. 13, 2182-2191.
- Okutani, Y. et al. (2001) Oncogene 20, 6643-6650.
- Demoulin, J.B. et al. (1999) J. Biol. Chem. 274, 25855-25861.
- Dentelli, P. et al. (1999) J. Immunol. 163, 2151-2159.
- Meinke, A. et al. (1996) Mol. Cell. Biol. 16, 6937-6944.
- Fata, J. E. et al. (2000) Cell 103, 41-50. Applications: IHC-P (paraffin).
- Kapur, R. and Zhang, L. (2000) J. Biol. Chem. 276, 1099-1106. Applications: Western Blotting.
- Siren, A. L. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4044-4049. Applications: Western Blotting.
- Reuther, G. W. et al. (2000) Mol. Cell. Biol. 20, 8655-8666. Applications: Western Blotting.
- Bergonzini, V. et al. (2004) Oncogene 23, 7701-7711. Applications: Western Blotting.
- Brizzi, M. F. et al. (2002) J. Clin. Invest. 109 (1), 111. Applications: Western Blotting, IC-IF, IHC-P (paraffin).
- Hutt, J. A. et al. (2000) J. Biol. Chem. 275, 29123-29131. Applications: Western Blotting.
- Weisberg, E. et al. (2011) PLoS One 6, e25351. Applications: Western Blotting.
- Pulikkan, J.A. et al. (2012) Blood , . Applications: Western Blotting.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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