Immunofluorescence analysis of Phospho-c-Kit (Tyr823) Polyclonal Antibody was performed using 70% confluent log phase Jurkat cells treated with 250 ng/ml SCF for 5 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-c-Kit (Tyr823) Rabbit Polyclonal Antibody (44498G) at 1:250 dilution 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 (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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the untreated control cells with no signal. Panel f shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Avian, Human|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human c-Kit that contains tyrosine 823. The sequence is conserved in cow and dog.|
|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 *|
|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.
|Western Blot (WB)||See 2 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.
CADM1 controls actin cytoskeleton assembly and regulates extracellular matrix adhesion in human mast cells.
44-498G was used in western blot to study the role of CADM1 in cytoskeleton assembly and regulates extracellular matrix adhesion in human mast cells
|Moiseeva EP,Straatman KR,Leyland ML,Bradding P||PloS one (9:null)||2014|
|Mouse||Not Cited||Lyn contributes to regulation of multiple Kit-dependent signaling pathways in murine bone marrow mast cells.||Shivakrupa R,Linnekin D||Cellular signalling (17:103)||2005|