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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; 100ng/ml for 10 min), and NK-92 cells, untreated or treated with Human Interleukin-2 #8907 (hIL-2; 100ng/ml for 10 min), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb (upper) or total Stat5 (3H7) Rabbit mAb #9358 (lower).Learn more about how we get our images
Western blot analysis of extracts from various cell lines using the Stat5 Antibody.Learn more about how we get our images
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.Learn more about how we get our images
Flow cytometric analysis of TF-1 cells, untreated (blue) or GM-CSF treated (green), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb.Learn more about how we get our images
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 10 μl of Stat5 Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Mouse CIS Intron 1 Primers #5131, mouse SOCS3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.Learn more about how we get our images
Gallery: PhosphoPlus® Stat5 (Tyr694) Antibody Kit #9350
|Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb 4322||100 µl||
||H M||90||Rabbit IgG|
|Stat5 Antibody 9363||100 µl||
||H M R Mk||90||Rabbit|
|Stat5 Control Cell Extracts 9353||100 µl||
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
|Anti-biotin, HRP-linked Antibody 7075||100 µl||
|Biotinylated Protein Ladder Detection Pack 7727||100 µl||
|20X LumiGLO® Reagent and 20X Peroxide 7003||5 ml each||
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.
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.
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.
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).
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PhosphoPlus is a trademark of Cell Signaling Technology, Inc. LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.