Immunofluorescence analysis of Phospho-c-Kit / CD117 pTyr721 Antibody was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-c-Kit / CD117 pTyr721 Antibody(44494G) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 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 594 Phalloidin (A12381). Panel d is a merged image showing membranous localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human c-Kit that contains tyrosine 721.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Western Blot (WB)||1:500-1:2000|
* 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.
|Western Blot (WB)||See 1 publications below|
c-Kit, also known as CD117 and stem cell factor receptor, is a 145 kDa transmembrane tyrosine kinase encoded by the c-Kit proto-oncogene. c-Kit acts to regulate a variety of biological responses including cell proliferation, apoptosis, chemotaxis and adhesion. Ligand binding to the extracellular domain leads to autophosphorylation on several tyrosine residues within the cytoplasmic domain, and activation. c-Kit mutations correlate with tumor growth and progression in a variety of cancers including mast cell disease, gastrointestinal stromal tumor, acute myeloid leukemia, Ewing sarcoma, and lung cancer. Phosphorylation at tyrosine 703 of c-Kit allows binding of Grb2 and activation of the Ras-Raf-ERK1 and 2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||Sorafenib inhibits imatinib-resistant KIT and platelet-derived growth factor receptor beta gatekeeper mutants.||Guida T,Anaganti S,Provitera L,Gedrich R,Sullivan E,Wilhelm SM,Santoro M,Carlomagno F||Clinical cancer research : an official journal of the American Association for Cancer Research (13:3363)||2007|