Cell Signaling Technology

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. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Stat5 (Tyr694) (14H2) Mouse mAb detects endogenous levels of Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699. It does not cross-react with 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 Tyr694 of mouse Stat5a.

Western Blotting

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 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).

  1. Gouilleux, F. et al. (1994) EMBO J. 13, 4361-4369.
  2. Wakao, H. et al. (1994) EMBO J. 13, 2182-2191.
  3. Okutani, Y. et al. (2001) Oncogene 20, 6643-6650.
  4. Demoulin, J.B. et al. (1999) J. Biol. Chem. 274, 25855-25861.
  5. Dentelli, P. et al. (1999) J. Immunol. 163, 2151-2159.
  6. Meinke, A. et al. (1996) Mol. Cell. Biol. 16, 6937-6944.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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