Equine IL-1 beta /IL-1F2 Antibody

  
  • Species Reactivity
    Equine
  • Specificity
    Detects equine IL‑1 beta /IL‑1F2 in direct ELISAs and Western blots. In direct ELISAs, approximately 100% cross‑reactivity with recombinant canine IL-1 beta, recombinant feline IL-1 beta, and recombinant human IL-1 beta  is observed, 40% cross-reactivity with recombinant porcine IL-1 beta, recombinant mouse IL-1 beta, and recombinant rat IL-1 beta is observed, and 10% cross-reactivity with recombinant cotton rat IL-1 beta and is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant equine IL‑1 beta /IL‑1F2
    Ala116-Ala268 (Glu179Gly, Met188Thr, Thr194Ile, Ser245Lys, Arg256Gln)
    Accession # Q28386
  • 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 Equine IL‑1 beta /IL‑1F2 (Catalog # 3340-EL)
  • Immunocytochemistry
    20-35 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑1 beta /IL‑1F2-induced proliferation in the D10.G4.1 mouse helper T cell line. Symons, J. A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 272. The Neutralization Dose (ND50) is typically 0.05-0.25 µg/mL in the presence of 100 pg/mL Recombinant Equine IL‑1 beta /IL‑1F2 and 1.25 µg/mL concanavalin A.
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‑1 beta /IL‑1F2 and Neutral­ization by Equine IL‑1 beta /IL‑1F2 Antibody.
Recombinant Equine IL‑1 beta /
IL‑1F2 (Catalog # 3340‑EL) stimulates proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Equine IL‑1 beta /IL‑1F2 (100 pg/mL) is neutralized (green line) by increasing concentrations of Goat Anti‑Equine IL‑1 beta /IL‑1F2 Antigen Affinity‑purified Poly­clonal Antibody (Catalog # AF3340). The ND50 is typically 0.05‑0.25 µg/mL in the presence of concanavalin A (1.25 µg/mL).
Immunocytochemistry
IL‑1 beta /IL‑1F2 in Equine PBMCs. IL‑1 beta /IL‑1F2 was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Equine IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3340) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) 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-1 beta/IL-1F2
IL-1 is a name that designates two pleiotropic cytokines, IL-1 alpha (IL-1F1) and IL-1 beta (IL-1F2), which are the products of distinct genes. IL-1 alpha and IL-1 beta are structurally related polypeptides that share approximately 27% amino acid (aa) identity in equine. Both proteins are produced by a wide variety of cells in response to inflammatory agents, infections, or microbial endotoxins. While IL-1 alpha and IL-1 beta are regulated independently, they bind to the same receptor and exert identical biological effects. IL-1 RI binds directly to IL-1 alpha or IL-1 beta and then associates with IL-1 R accessory protein (IL-1 R3/IL-1 R AcP) to form a high-affinity receptor complex that is competent for signal transduction. IL-1 RII has high affinity for IL-1 beta but functions as a decoy receptor and negative regulator of IL-1 beta activity. IL-1ra functions as a competitive antagonist by preventing IL-1 alpha and IL-1 beta from interacting with IL-1 RI (1-4). The equine IL-1 beta cDNA encodes a 268 aa precursor. A 115 aa propeptide is cleaved intracellularly by the cysteine protease IL-1 beta -converting enzyme (Caspase-1/ICE) to generate the active cytokine (5-7). An alternatively spliced form of equine IL-1 beta has a deletion which encompasses the Caspase-1 cleavage site and potentially results in a membrane-associated form (8). The 17 kDa mature equine IL-1 beta shares 65%-75% aa sequence identity with canine, cotton rat, feline, human, mouse, porcine, rat, and rhesus IL-1 beta.
  • References:
    1. Allan, S.M. et al. (2005) Nat. Rev. Immunol. 5:629.
    2. Boraschi, D. and A. Tagliabue (2006) Vitam. Horm. 74:229.
    3. Kornman, K.S. (2006) Am. J. Clin. Nutr. 83:475S.
    4. Isoda, K. and F. Ohsuzu (2006) J. Atheroscler. Thromb. 13:21.
    5. Kato, H. et al. (1997) Vet. Immunol. Immunopathol. 48:221.
    6. Howard, R.D. et al. (1998) Am. J. Vet. Res. 59:704.
    7. Martinon, F. and J. Tschopp (2007) Cell Death Differ. 14:10.
    8. Kato, H. et al. (1996) Gene 177:11.
  • Long Name:
    Interleukin 1 beta
  • Entrez Gene IDs:
    3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
  • Alternate Names:
    catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta
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