Detection of Human Phospho-CD117/ c‑kit (Y730) by Western Blot.
Western blot shows lysates of MO7e human megakaryocytic leukemic cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human SCF/c‑kit Ligand (Catalog # 255-SC) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-CD117/c-kit (Y730) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3527), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-CD117/c-kit (Y730) at approximately 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Stem cell factor receptor (CD117, the gene product of the c-kit proto-oncogene) and its ligand, stem cell factor (also named c-kit ligand, mast cell growth factor), play essential roles in gametogenesis, melanogenesis and hematopoiesis. The human stem cell factor receptor cDNA encodes a 972 amino acid (aa) residue precursor membrane protein with a 25 aa residue signal peptide (experimentally determined), a 495 aa residue extracellular domain, a 23 aa residue transmembrane segment and a 429 aa residue cytoplasmic domain. Stem cell factor receptor is a member of the type III subfamily of receptor tyrosine kinases (RTK) that also includes the receptors for M-CSF, Flt-3, PDGF and VEGF. All class III RTKs are characterized by the presence of five immunoglobulin-like domains in their extracellular region and a split kinase domain in their intracellular region. SCF binding induces receptor homodimerization and signal transduction. SCF receptor is expressed in hematopoietic progenitor cells, normal B- and T-lymphocyte progenitor cells, mast cells, germ cells, melanocytes, neurons, glial cells, placenta, kidney, lung and gut. In addition, SCF receptor expression has also been reported in a number of human tumor cell lines. SCF receptor can be proteolytically cleaved from the cell surface and high levels of soluble SCF receptor has been detected in cell conditioned medium and human plasma. Recombinant soluble SCF receptor binds SCF with high affinity and is a potent SCF antagonist.
Broudy, V. (1997) Blood 90:1345.
Vliagoftis, H. et al. (1997) J. Allergy Clin. Immunol. 100:435.
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