Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

PhosphoPlus® Stat5 (Tyr694) Antibody Kit #9350

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Stat5 Antibody # 9363 100 microliters W IP H M R 90 Rabbit
Phospho-Stat5 (Tyr694) Antibody # 9351 100 microliters W F H M (R) (B) 90 Rabbit
Stat5 Control Cell Extracts # 9353 40 microliters Human
Anti-rabbit IgG, HRP-linked Antibody # 7074 50 microliters Goat
Anti-biotin, HRP-linked Antibody # 7075 100 microliters Goat
20X LumiGLO® Reagent and 20X Peroxide # 7003 5 milliliters
Biotinylated Protein Ladder Detection Pack # 7727 100 microliters

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine

Specificity / Sensitivity

Phospho-Stat5 (Tyr694) Antibody detects endogenous levels of Stat5alpha and Stat5beta only when phosphorylated at Tyr694. This antibody does not cross-react with the corresponding phospho-tyrosine residues of other Stat proteins, but does cross-react with EGFR. Stat5 Antibody detects endogenous levels of total Stat5alpha and Stat5beta.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, K562 and TF-1 cells, using the Stat5 Antibody.

IC-ABC

IC-ABC

Phospho-Stat5 (Tyr694) Antibody #9351 allows in situ detection of IFN-alpha induced Stat5 nuclear translocation in HeLa cells.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K562 cells, untreated (blue) or STI571-treated (green), using Phospho-Stat5 (Tyr694) Antibody #9351 compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Immunofluorescent staining of HeLa cells, untreated (left) or IFN-alpha-treated (100 ng/ml for 15 minutes), using Phospho-Stat5 (Tyr694) Antibody #9351 and anti-rabbit Alexa Fluor®4 88 as the secondary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues around Tyr694 of mouse Stat5 (Phospho-Stat5 (Tyr694) Antibody) or a synthetic peptide (KLH-coupled) corresponding to an amino-terminal region of Stat5. Antibodies were purified by protein A and peptide affinity chromatography.

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).

  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

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.

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