Immunofluorescence analysis of Phospho-STAT5 pTyr694 was done on 70% confluent log phase HeLa cells treated with 5uM of TNF-alpha for 30 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-STAT5 pTyr694 Rabbit Polyclonal antibody (44390G) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is untreated cell with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human STAT5a that contains tyrosine 694. The sequence is conserved in mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-5 µg|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The protein encoded by this gene is a member of the STAT family of transcription factors. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein is activated by, and mediates the responses of many cell ligands, such as IL2, IL3, IL7 GM-CSF, erythropoietin, thrombopoietin, and different growth hormones. Activation of this protein in myeloma and lymphoma associated with a TEL/JAK2 gene fusion is independent of cell stimulus and has been shown to be essential for the tumorigenesis. The mouse counterpart of this gene is found to induce the expression of BCL2L1/BCL-X(L), which suggests the antiapoptotic function of this gene in cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The Placental Variant of Human Growth Hormone Reduces Maternal Insulin Sensitivity in a Dose-Dependent Manner in C57BL/6J Mice.
44-390G was used in western blot to test the effect of maternal GH variant administration on maternal and fetal growth and metabolic outcomes during pregnancy
|Liao S,Vickers MH,Stanley JL,Ponnampalam AP,Baker PN,Perry JK||Endocrinology (157:1175)||2016|
Urokinase-type plasminogen activator receptor signaling is critical in nasopharyngeal carcinoma cell growth and metastasis.
44-390G was used in western blot to show that uPAR regulates nasopharyngeal carcinoma NPC progression.
|Bao YN,Cao X,Luo DH,Sun R,Peng LX,Wang L,Yan YP,Zheng LS,Xie P,Cao Y,Liang YY,Zheng FJ,Huang BJ,Xiang YQ,Lv X,Chen QY,Chen MY,Huang PY,Guo L,Mai HQ,Guo X,Zeng YX,Qian CN||Cell cycle (Georgetown, Tex.) (13:1958)||2014|
|Mouse||Not Cited||Germline CBL mutations cause developmental abnormalities and predispose to juvenile myelomonocytic leukemia.||Niemeyer CM,Kang MW,Shin DH,Furlan I,Erlacher M,Bunin NJ,Bunda S,Finklestein JZ,Sakamoto KM,Gorr TA,Mehta P,Schmid I,Kropshofer G,Corbacioglu S,Lang PJ,Klein C,Schlegel PG,Heinzmann A,Schneider M,Starý J,van den Heuvel-Eibrink MM,Hasle H,Locatelli F,Sakai D,Archambeault S,Chen L,Russell RC,Sybingco SS,Ohh M,Braun BS,Flotho C,Loh ML||Nature genetics (42:794)||2010|