Canine SCF/c-kit Ligand Antibody AF2278: R&D Systems

Canine SCF/c-kit Ligand Antibody

  
  • Species Reactivity
    Canine
  • Specificity
    Detects canine SCF/c‑kit Ligand in ELISAs and Western blots. In sandwich immunoassays, less than 20% cross-reactivity with recombinant feline SCF is observed, less than 8% cross-reactivity with recombinant human SCF is observed, and less than 0.5% cross-reactivity with recombinant mouse SCF is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine SCF/c‑kit Ligand
    Lys26-Ala190
    Accession # Q06220
  • 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
    Recombinant Canine SCF/c-kit Ligand (Catalog # 2278-SC)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Canine SCF/c-kit Ligand Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Canine SCF/c‑kit Ligand Antibody (Catalog # AF2278)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Canine SCF/c‑kit Ligand Biotinylated Antibody (Catalog # BAF2278)
  • ELISA Standard
     
    Recombinant Canine SCF Protein (Catalog # 2278-SC)
  • 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 0.1-0.3 µg/mL in the presence of 5 ng/mL Recombinant Canine 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 Canine SCF/c‑kit Ligand Antibody. Recombinant Canine SCF/c‑kit Ligand (Catalog # 2278-SC) stimulates pro­liferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Canine SCF/c‑kit Ligand (5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine SCF/
c-kit Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2278). The ND50 is typically 0.1‑0.3 µg/mL.
Immunocytochemistry
SCF/c‑kit Ligand in Canine PBMCs. SCF/c‑kit Ligand was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Canine SCF/c‑kit Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2278) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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: SCF/c-kit Ligand

Canine SCF (stem cell factor; also known as c‑kit ligand) is a type I transmembrane (TM) glycoprotein that plays an important role in a number of fetal and adult developmental processes (1‑4). It is synthesized as a 274 amino acid (aa) precursor that contains a 25 aa signal sequence, a 190 aa extracellular region, a 23 aa TM segment and a 36 aa cytoplasmic tail (5). Within the extracellular region there are four potential N‑linked glycosylation sites, two intrachain disulfide bonds, and four alpha ‑helices. Although the predicted molecular weight is 19 kDa, the native molecule is anywhere from 28‑40 kDa in size and reflects both N‑ and O‑linked glycosylation (1). Glycosylation is not necessary for bioactivity (6). The transmembrane form of SCF can be cleaved proteolytically, generating a 165 aa soluble form. Circulating SCF exists as both a monomer and nondisulfide‑linked homodimer, with monomer predominating (50% to 75%) (6). Both the soluble and TM forms have bioactivity. Their principal targets may be different, however (7). A second, alternate splice short form of SCF has been identified in other species (1). It is membrane bound but lacks the proteolytic cleavage site found in the long form. Thus, it cannot give rise to a soluble molecule. No such isoform has been reported for canine, but it could be assumed to exist. The ratio of long form to short form varies from tissue to tissue (1). Soluble canine SCF shares 88%, 93%, 86%, 83%, 76%, 76%, 86% and 88% aa sequence identity with porcine, feline, bovine, human, mouse, rat, goat and equine SCF, respectively. Cells known to express SCF include endothelial cells, fibroblasts and keratinocytes (1).

  • References:
    1. Broudy, V.C. (1997) Blood 90:1345.
    2. Nakagawa, S. and T. Kitoh (2000) Curr. Opin. Hematol. 7:133.
    3. Yoshida, H. et al. (2001) J. Invest. Dermatol. Symp. Proc. 6:1.
    4. Kang, J. and S.D. Der (2004) Curr. Opin. Immunol. 16:180.
    5. Dunham, S.P. and D.E. Onions (1996) DNA Seq. 6:233.
    6. Hsu, Y-R. et al. (1997) J. Biol. Chem. 272:6406.
    7. Kapur, R. et al. (1998) Blood 91:879.
  • Long Name:
    Stem Cell Factor
  • Entrez Gene IDs:
    4254 (Human); 17311 (Mouse); 60427 (Rat); 403507 (Canine); 493937 (Feline)
  • Alternate Names:
    c-kit Ligand; DKFZp686F2250; familial progressive hyperpigmentation 2; FPH2; KIT ligand; Kitl; KITLG; KL-1; Mast cell growth factor; MGF; MGFSHEP7; SCF; SCFStem cell factor; SFc-Kit ligand; SLF; steel factor
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