PhosphoPlus® Stat5 (Tyr694) Antibody Kit #9350
Western blot analysis of extracts from UT-7 cells, untreated or treated with erythropoietin (EPO; 3 units/ml for 5 min), TF-1 cells, untreated or treated with Human Granulocyte Macrophage Colony Stimulating Factor #8922 (hGM-CSF; 100 ng/ml for 10 min), and NK-92 cells, untreated or treated with Human Interleukin-2 #8907 (hIL-2; 100 ng/ml for 10 min), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb.
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Product Information
Kit Usage Information
Protocols
Product Description
The PhosphoPlus® Stat5 (Tyr694) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of Stat5a at Tyr694 and Stat5b at Tyr699.
| MW (kDa) | 90 |
Specificity / Sensitivity
Phospho-Stat5 (Tyr694) (D47E7) XPTM Rabbit mAb detects endogenous levels of Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699. Stat5 (3H7) Rabbit mAb detects endogenous levels of total Stat5α and ß protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr694 of human Stat5a protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to an amino-terminal region within 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 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).
Pathways
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