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Peptide Competition: Extracts prepared from M07e cells left untreated (1) or treated (2-5) with SCF were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 µg/ml c-Kit [pY703] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phospho-tyrosine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab"e;)2 anti-rabbit IgG alkaline phosphatase (Cat. no. ALI4405) and signals were detected using the Tropix WesternStar™ method. The data show that only the peptide corresponding to c-Kit [pY703] blocks the antibody signal, thereby demonstrating the specificity of the antibody.
|Tested species reactivity||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 703.|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||0.1-1.0 ug/ml|
* 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&2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||1:500||The identification and characterisation of novel KIT transcripts in aggressive mast cell malignancies and normal CD34+ cells.||Ozer O,Zhao YD,Ostler KR,Akin C,Anastasi J,Vardiman JW,Godley LA||Leukemia & lymphoma (49:1567)||2008|
|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|
C-Kit; CD117; ckit; Mast/stem cell growth factor receptor Kit; p145 c-kit; PBT; Piebald trait protein; Proto-oncogene c-Kit; proto-oncogene tyrosine-protein kinase Kit; SCFR; soluble KIT variant 1; Tyrosine-protein kinase Kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein
C-Kit; CD117; KIT; PBT; SCFR