hh Human CD117/c-kit Antibody AF332: R&D Systems

Human CD117/c-kit Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human and mouse CD117/c-kit in direct ELISAs and Western blots. In Western blots, approximately 2% cross-reactivity with recombinant human PDGF sR alpha is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD117/c-kit
    Gln26-Thr520
    Accession # P10721
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Flow Cytometry
    2.5 µg/106 cells
    TF‑1 human erythroleukemic cell line
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Neutralization
    Measured by its ability to neutralize SCF/c‑kit Ligand-induced proliferation in the TF‑1 human erythroleukemic cell line. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323.The Neutralization Dose (ND50) is typically 0.06-0.36 μg/mL in the presence of 20 ng/mL Recombinant Human SCF/c‑kit Ligand.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cell Proliferation Induced by SCF/c‑kit Ligand and Neutralization by Human CD117/c-kit Antibody. Recombinant Human SCF/c‑kit Ligand (Catalog # 255-SC) induces cell proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Under these conditions, proliferation elicited by
SCF/c‑kit Ligand is neutralized (green line) by increasing concentrations of Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332). The ND50 is typically 0.06-0.36 μg/mL in the presence of 20 ng/mL of Recombinant Human
SCF/c‑kit Ligand.
Immunohistochemistry
CD117/c-kit in Human Brain. CD117/c-kit was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human CD117/c-kit by Western Blot. Western blot shows lysates of MO7e human megakaryocytic leukemic cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD117/c-kit at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human CD117/c-kit by Simple WesternTM. Simple Western lane view shows lysates of MO7e human megakaryocytic leukemic cell line, loaded at 0.2 mg/mL. A specific band was detected for CD117/c-kit at approximately 172 kDa (as indicated) using 5 µg/mL of Goat Anti-Human CD117/c-kit Antigen Affinity-purified Polyclonal Antibody (Catalog # AF332) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.        
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: CD117/c-kit

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.

  • References:
    1. Broudy, V. (1997) Blood 90:1345.
    2. Vliagoftis, H. et al. (1997) J. Allergy Clin. Immunol. 100:435.
  • Long Name:
    Stem Cell Factor Receptor
  • Entrez Gene IDs:
    3815 (Human); 16590 (Mouse); 64030 (Rat)
  • Alternate Names:
    CD117 antigen; CD117; ckit; c-kit; EC 2.7.10; EC 2.7.10.1; KIT; PBT; piebald trait; Proto-oncogene c-Kit; proto-oncogene tyrosine-protein kinase Kit; SCF R; SCFR; SCFRmast/stem cell growth factor receptor; 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Regulation of human skin pigmentation in situ by repetitive UV exposure: molecular characterization of responses to UVA and/or UVB.
    Authors: Choi W, Miyamura Y, Wolber R, Smuda C, Reinhold W, Liu H, Kolbe L, Hearing VJ
    J. Invest. Dermatol., 2010;130(6):1685-96.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  2. Chemotherapy-induced thrombocytopenia derives from the selective death of megakaryocyte progenitors and can be rescued by stem cell factor.
    Authors: Zeuner A, Signore M, Martinetti D, Bartucci M, Peschle C, De Maria R
    Cancer Res., 2007;67(10):4767-73.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  3. Granulocyte colony-stimulating factor induces the release in the bone marrow of proteases that cleave c-KIT receptor (CD117) from the surface of hematopoietic progenitor cells.
    Authors: Levesque JP, Hendy J, Winkler IG, Takamatsu Y, Simmons PJ
    Exp. Hematol., 2003;31(2):109-17.
    Species: Human
    Sample Type: Recombinant Protein
    Application: WB
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Normal Goat IgG Control

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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

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Goat IgG Horseradish Peroxidase-conjugated Antibody

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