Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3939

Applications Reactivity Sensitivity MW (kDa) Isotype
F H M (R) (Mk) (B) Endogenous 90 Rabbit IgG

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine
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) (C71E5) Rabbit mAb (Alexa Fluor® 488) detects endogenous levels of Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr694 of Stat5a. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of TF-1 cells, untreated (blue) or treated with GM-CSF (green), using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 488 Conjugate).

Description

This Cell Signaling Technology (CST) antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry in human cells. CST expects that Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 488) will display the same species cross-reactivity as the unconjugated antibody (Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb #9314).

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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