Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb #9359

Applications Reactivity MW (kDa) Source Isotype
W IP IHC-P IF-IC F H M (R) (Mk) (B) 90 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb detects endogenous levels of Stat5α and Stat5β proteins when phosphorylated at Tyr694.

Source / Purification

Rabbit monoclonal antibodies were prepared from spleens of rabbits immunized with a synthetic peptide (KLH-coupled) corresponding to residues surrounding Tyr694 of Stat5.

Western Blotting

Western Blotting

Western blot analysis of extracts from K562 cells, untreated or treated with Gleevec, using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb (top) and total Stat5 (3H7) Rabbit mAb #9358 (bottom).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue) using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or IFNα-treated (right) and labeled with Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red).

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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