Product Pathways - Jak/Stat Pathway
Phospho-Stat5 (Tyr694) (14H2) Mouse mAb #9356
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP | H M (R) | Endogenous | 90 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Protocols
Specificity / Sensitivity
Phospho-Stat5 (Tyr694) (14H2) Mouse mAb detects endogenous levels of Stat5-alpha and Stat5-beta only when phosphorylated at tyrosine 694. It does not cross-react with the corresponding phospho-tyrosines of other Stat proteins.
Source / Purification
Monoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr694 of mouse Stat5.
Western Blotting
Western blot analysis of extracts from K562 and TF-1 cells, untreated, STI571-treated or GM-CSF-treated as indicated, using Phospho-Stat5 (Tyr694) (14H2) Mouse mAb (upper) or Stat5 Antibody (lower).
Background
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 has been found to be 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). Stat5α and β are independently regulated and activated in various cell types. For instance, both isoforms are activated in response to IFN-α in B cells, but only Stat5α is phosphorylated in response to IFN-α in HeLa cells (6).
- Gouilleux, F. et al. (1994) EMBO J. 13, 4361-4369.
- 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.
Application References
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Companion Products
- 9351 Phospho-Stat5 (Tyr694) Antibody
- 9914 Phospho-Stat Antibody Sampler Kit
- 7076 Anti-mouse 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
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
