Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #9365

Applications Reactivity MW (kDa) Isotype
F H M (R) (Mk) (B) 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® 647 Conjugate) 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® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

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® 647 Conjugate).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. CST expects that Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) 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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063) from Epitomics, Inc.

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

Products