Porcine IL-5 Antibody AF3137: R&D Systems

Porcine IL-5 Antibody

  
  • Species Reactivity
    Porcine
  • Specificity
    Detects porcine IL-5 in direct ELISAs and Western blots. In direct ELISAs, less than 50% cross‑reactivity with recombinant canine IL‑5 and recombinant feline IL-5 is observed and and less than 30% cross-reactivity with recombinant human IL-5, recombinant mouse IL-5, and recombinant rat IL-5 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant porcine IL-5
    Ile20-Ser134
    Accession # Q9MYM5
  • 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 Porcine IL‑5 (Catalog # 3137-PL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑5-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 2-8 µg/mL in the presence of 25 ng/mL Recombinant Porcine IL‑5.
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 IL‑5 and Neutralization by Porcine IL‑5 Antibody. Recombinant Porcine IL‑5 (Catalog # 3137-PL) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Porcine IL‑5 (25 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Porcine IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3137). The ND50 is typically
2‑8 µg/mL.
Immunocytochemistry
IL‑5 in Porcine PBMCs.
IL‑5 was detected in immersion fixed porcine peripheral blood mononuclear cells treated with calcium ionomycin and PMA using Goat Anti-Porcine IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3137) 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 cytoplasm. 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: IL-5

Interleukin-5 (IL-5) is a secreted disulfide-linked homodimeric glycoprotein that belongs to the alpha -helical group of cytokines that includes IL-3, IL-5 and GM-CSF (1‑3). IL-5 is primarily produced by CD4+ Th2 cells, but eosinophils and mast cells also produce IL-5. Porcine IL-5 is synthesized as a 134 amino acid (aa) precursor that contains a 19 aa signal sequence and a 115 aa mature segment (5). Four alpha -helices and two cysteines that form interchain disulfide bonds with a second, antiparallel IL-5 molecule are conserved among species (3 - 5). Monomeric IL-5 is a predicted 14 kDa protein but usage of N-linked glycosylation sites may increase its molecular weight (5). Mature porcine IL-5 shares 90%, 88%, 86%, 85%, 84%, 66%, 68%, 63%, 63% and 59% aa sequence identity with mature bovine, sheep, cat, equine, canine, human, guinea pig, cotton rat, murine and rat IL-5, respectively. Recombinant porcine IL-5 induced proliferation in the human TF-1 cell line (5). The receptor for human IL-5 consists of a 60 kDa ligand-binding subunit (IL-5 R alpha ) and a 120 kDa signal-transducing subunit ( beta c). It is suggested that dimeric IL-5 binding to IL-5 R alpha recruits beta c, which subsequently covalently links with IL-5 R alpha. Two receptor complexes then associate to form the physiologic IL-5 receptor (6, 7). IL-5 binds proteoglycans, potentially enhancing its activity (8). Following receptor binding, IL-5 promotes the maturation, activation and migration of eosinophils, as demonstrated during asthmatic eosinophilic lung inflammation (1‑3). It also mobilizes eosinophils and CD34+ progenitors from marrow. It also enhances Ig release from B cells and contributes to IL-4 production. Finally, it primes basophils for histamine and leukotriene release (1, 2, 9).

  • References:
    1. Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
    2. Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
    3. Zabeau, L. et al. (2003) Curr. Drug Targets Inflamm. Allergy 2:319.
    4. Mertens, B. et al. (1996) Gene 176:273.
    5. Sylvin, H. et al. (2000) Immunogenetics 51:59.
    6. Bagley, C.J. et al. (1997) Blood 89:1471.
    7. Ishino, T. et al. (2005) J. Biol. Chem. 280:22951.
    8. Lipscombe, R. et al. (1998) J. Leukocyte Biol. 63:342.
    9. Mattes, J. and P.S. Foster (2003) Curr. Drug Targets Inflamm. Allergy 2:169.
  • Long Name:
    Interleukin 5
  • Entrez Gene IDs:
    3567 (Human); 16191 (Mouse); 24497 (Rat); 397409 (Porcine); 280825 (Bovine); 403790 (Canine); 493803 (Feline)
  • Alternate Names:
    BCDF mu; B-cell differentiation factor I; BCGFII; EDF; Eo-CSF; Eosinophil differentiation factor; IL5; IL-5; IL-5T-cell replacing factor; interleukin 5 (colony-stimulating factor, eosinophil); interleukin-5; TRF; TRFB cell differentiation factor I
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