hh Human/Rhesus Macaque IL-18/IL-1F4 Antibody AF2548: R&D Systems

Human/Rhesus Macaque IL-18/IL-1F4 Antibody

  
  • Species Reactivity
    Human, Rhesus Macaque
  • Specificity
    Detects rhesus macaque IL‑18/IL-1F4 in direct ELISAs and Western blots. In Western blots, approximately 10% cross-reactivity with recombinant mouse IL-18, recombinant rat IL-18, and recombinant porcine IL-18 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rhesus macaque IL‑18/IL-1F4
    Tyr37-Asp193
    Accession # AAK13416
  • 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 Rhesus Macaque IL‑18/IL‑1F4 (Catalog # 2548-RM)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑18/IL‑1F4-induced IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line. Novick, D. et al. (1999) Immunity 10(1):127. The Neutralization Dose (ND50) is typically 0.4‑1.2 µg/mL in the presence of 10 ng/mL Recombinant Rhesus Macaque IL‑18/IL‑1F4 and 20 ng/mL Recombinant Human TNF‑ alpha.
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
IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Primate IL‑18/IL‑1F4 Antibody. In the presence of Recombinant Human TNF‑ alpha (20 ng/mL, Catalog # 210-TA), Recombinant Rhesus Macaque IL‑18/IL‑1F4 (Catalog # 2548-RM) stimulates IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line in a dose-dependent manner (orange line), as measured by the Human IFN‑ gamma Quantikine ELISA Kit (Catalog # DIF50). Under these conditions, IFN‑ gamma secretion elicited by Recombinant Rhesus Macaque IL‑18/IL‑1F4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Rhesus Macaque IL‑18/IL‑1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548). The ND50 is typically 0.4-1.2 µg/mL.
Immunocytochemistry
IL‑18/IL‑1F4 in Human PBMCs. IL‑18/IL‑1F4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human/Rhesus Macaque IL‑18/IL‑1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) at 5 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 (green). 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-18/IL-1F4
Interleukin-18 (IL-18), also known as IL-1F4 and IFN-gamma inducing factor (IGIF), is a member of the IL-1 family of cytokines and is a key molecule in the innate immune response (1). Rhesus IL-18 is synthesized as a 24 kDa proprotein that contains a 36 amino acid (aa) propeptide and a 157 aa mature region (2). Under inflammatory conditions, the propeptide is cleaved by Caspase-1 in the cytoplasm to liberate the mature nonglycosylated 18 kDa monomeric IL-18 (3, 4). Mature rhesus IL-18 shares 96% aa sequence identity with human IL-18 and 60-76% with mouse, rat, canine, feline, and porcine IL-18. IL-18 is secreted by a variety of cell types including macrophages, dendritic cells, and epithelial cells (1, 5). Circulating mature IL-18 is sequestered by soluble IL-18 binding proteins (IL-18 BP) that inhibit IL-18 bioactivity (6). IL-18 interacts with the widely expressed IL-18 R alpha which then recruits the signaling subunit IL-18 R beta (7, 8). The IL-1 family member IL-1F7 also binds to IL-18 R alpha but does not recruit IL-18 R beta or induce signaling (9). IL-1F7 binds IL-18 BP and enhances its neutralizing effect on IL-18 activity (9). IL-18 synergizes with other cytokines to activate NK, Th1, and Th17 cells and to increase the production of IFN-gamma (1, 5, 10-12). IL-18 can also promote Th2 cytokine release which reduces the effectiveness of antiviral responses (13, 14). Increased levels of active IL-18 contribute to the severity of autoimmunity and hypertension, while deficiency of IL-18 results in symptoms of metabolic syndrome (1, 5, 15, 16). In cancer, IL-18 stimulates Th1 and NK cells to target tumor cells, but it can also promote angiogenesis, metastasis, and tumor cell immune evasion (11).
  • References:
    1. Arend, W.P. et al. (2008) Immunol. Rev. 223:20.
    2. Giavedoni, L.D. et al. (2001) J. Interferon Cytokine Res. 21:173.
    3. Ghayur, T. et al. (1997) Nature 386:619.
    4. Gu, Y. et al. (1997) Science 275:206.
    5. Boraschi, D. and C.A. Dinarello (2006) Eur. Cytokine Netw. 17:224.
    6. Novick, D. et al. (1999) Immunity 10:127.
    7. Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
    8. Born, T.L. et al. (1998) J. Biol. Chem. 273:29445.
    9. Bufler, P. et al. (2002) Proc. Natl. Acad. Sci. USA 99:13723.
    10. Takeda, K. et al. (1998) Immunity 8:383.
    11. Park, S. et al. (2007) Cell. Mol. Immunol. 4:329.
    12. Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
    13. Hoshino, T. et al. (2001) J. Immunol. 166:7014.
    14. Iannello, A. et al. (2009) AIDS Rev. 11:115.
    15. Rabkin, S.W. (2009) Nat. Clin. Pract. Cardiovasc. Med. 6:192.
    16. Netea, M.G. et al. (2006) Nat. Med. 12:650.
    17. Long Name:
      Interleukin 18
    18. Entrez Gene IDs:
      3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)
    19. Alternate Names:
      Iboctadekin; IFN-gamma-inducing factor; IGIF; IGIFIL-1 gamma; IL18; IL-18; IL-18MGC12320; IL-1F4; IL1F4iboctadekin; IL-1g; Interferon gamma-inducing factor; interleukin 18 (interferon-gamma-inducing factor); Interleukin-1 gamma; interleukin-18
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