Equine IL-1ra/IL-1F3 Antibody

  
  • Species Reactivity
    Equine
  • Specificity
    Detects equine IL‑1ra/IL‑1F3 in ELISAs and Western blots. In sandwich immunoassays, less than 20% cross-reactivity with recombinant mouse IL-1ra is observed, less than 5% cross-reactivity with recombinant rat IL-1ra is observed, less than 2% cross-reactivity with recombinant porcine IL-1ra is observed, less than 0.4% cross-reactivity with recombinant human IL-1ra is observed, and less than 0.2% cross-reactivity with recombinant equine IL-1beta is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant equine IL‑1ra/IL‑1F3
    His26-Gln177
    Accession # O18999
  • 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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Equine IL‑1ra/IL‑1F3 (Catalog # 2466-RA)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Equine IL-1ra/IL-1F3 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Equine IL‑1ra/IL‑1F3 Antibody (Catalog # AF2466)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Equine IL‑1ra/IL‑1F3 Biotinylated Antibody (Catalog # BAF2466)
  • ELISA Standard
     
    Recombinant Equine IL-1ra/IL-1F3 Protein (Catalog # 2466-RA)
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
Immunocytochemistry
IL‑1ra/IL‑1F3 in Equine PBMCs.

IL‑1ra/IL‑1F3 was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Equine IL‑1ra/IL‑1F3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2466) 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: IL-1ra/IL-1F3

Secreted equine IL-1 receptor antagonist (IL-1ra) is a presumably 22‑25 kDa glycoprotein produced by variety of cell types that antagonizes IL-1 activity (1‑3). It is a member of the IL-1 family of proteins that includes IL-1 alpha and IL-1 beta. Although there is little amino acid (aa) identity (<30%) among the three IL-1 family members, all molecules bind to the same receptors, all show a beta -trefoil structure, and all are believed to have evolved from a common ancestral gene (1‑4). Equine IL-1ra is synthesized as a 177 aa precursor that contains a 25 aa signal sequence plus a 152 aa mature region. There is one intrachain disulfide bond and one potential N-linked glycosylation site (3, 5, 6). Mature equine sIL-1ra is 78%, 78%, 80%, 82%, and 76% aa identical to mature mouse, human, porcine, canine and bovine IL‑1ra, respectively. In human, three non-secreted IL-1ra isoforms have also been identified. It is unknown if such an analogous situation exists in equine. Cells known to secrete IL-1ra include fibroblasts, vascular smooth muscle cells, intestinal columnar epithelium, chondrocytes, macrophages, mast cells, neutrophils and hepatocytes.

There are two type I transmembrane glycoprotein receptors for IL-1ra. The first is the bioactive 80 kDa type I IL-1 receptor (IL-1 RI), and the second is the inert (decoy) 65 kDa type II IL-1 receptor. IL-1ra binding to IL-1 RI competitively blocks IL-1 ( alpha or beta ) binding to the same receptor. This results in receptor ligation without activation (1, 7). The type II IL-1 receptor is inert, and any binding of IL-1ra not only fails to block co-existing IL-1 activity, but may actually potentiate it by removing an IL-1 antagonist. Functionally, all activities attributed to IL-1ra are explained by its role as a competitive inhibitor of IL-1 binding to IL-1 RI (1, 2, 8, 9).

  • References:
    1. Arend, W.P. et al. (1998) Annu. Rev. Immunol. 16:27.
    2. Roux-Lombard, P. (1998) Eur. Cytokine Netw. 9:565.
    3. Dayer-J-M. (2002) Clin. Exp. Rheumatol. 20(27):S14.
    4. Eisenberg, S.P. et al. (1991) Proc. Natl. Acad. Sci. USA 88:5232.
    5. Kato, H. et al. (1997) Vet. Immunol. Immunopathol. 56:221.
    6. Howard, R.D. et al. (1998) Am. J. Vet. Res. 59:712.
    7. Dinarello, C.A. (1997) Semin. Oncol. (Suppl 9):S9.
    8. Irikura, V.M. et al. (2002) J. Immunol. 169:393.
    9. Arend, W.P. and C. Gabay (2000) Arth. Res. 2:245.
  • Long Name:
    Interleukin 1 Receptor Antagonist
  • Entrez Gene IDs:
    3557 (Human); 16181 (Mouse); 60582 (Rat); 397499 (Porcine)
  • Alternate Names:
    DIRA; ICIL-1RA; IL1F3; IL-1F3; IL1ra; IL-1ra; IL-1ra3; IL1RN; IL-1RN; interleukin 1 receptor antagonist; IRAP; MVCD4
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