Feline IL-12/IL-23 p40 Antibody

(1 citations)   
  • Species Reactivity
    Feline
  • Specificity
    Detects feline IL‑12/IL‑23 p40 in ELISAs and Western blots. In sandwich immunoassays, approximately 65% cross‑reactivity with recombinant canine IL-12/23 p40 is observed, less than 3% cross-reactivity with recombinant mouse IL-12/23 p40 is observed, and less than 0.2% cross-reactivity with recombinant human IL‑12/23 p40 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant feline IL‑12/IL‑23 p40
    Ile23-Ser329 (Glu167Gly)
    Accession # O02744
  • 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 Feline IL‑12/IL‑23 p40 (Catalog # 2117-FL)
  • Immunocytochemistry
    10-25 µg/mL
    See below
    • Feline IL-12/IL-23 p40 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Feline IL‑12/IL‑23 p40 Antibody (Catalog # AF1954)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Feline IL‑12/IL‑23 p40 Biotinylated Antibody (Catalog # BAF1954)
  • ELISA Standard
     
    Recombinant Feline IL-12/IL-23 p40 Protein (Catalog # 2117-FL)
  • Neutralization
    Measured by its ability to neutralize IL‑12-induced proliferation in PHA-activated human peripheral blood mononuclear cells (PBMC). Yokota, T. et al. (1986) Proc. Natl. Acad. Sci. USA 83:5894. The Neutralization Dose (ND50) is typically 0.5-2.0 µg/mL in the presence of 25 ng/mL Recombinant Feline IL‑12.
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‑12 and Neutralization by Feline IL‑12/IL‑23 p40 Antibody. Recombinant Feline IL‑12 (Catalog # 1954‑FL) stimulates proliferation in PHA-activated human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Feline IL‑12 (25 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Feline
IL‑12/IL‑23 p40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1954). The ND50 is typically 0.5-2.0 µg/mL.
Immunocytochemistry
IL‑12/IL‑23 p40 in Feline PBMCs. IL‑12/IL‑23 p40 was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) stimulated with PMA and calcium ionomycin using Goat Anti-Feline IL‑12/IL‑23 p40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1954) 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-12/IL-23 p40

Interleukin 12 (IL-12) and IL-23 are secreted heterodimeric glycoproteins belonging to the IL-12 cytokine family. The two cytokines share a common p40 (40 kDa) subunit, which is disulfide-linked with the p35 (35 kDa) subunit in IL-12, and with the p19 (19 kDa) subunit in IL-23. Feline p40 is synthesized as a 329 amino acid (aa) precursor with a 22 aa signal sequence and a 307 aa mature region. It contains a 90 aa fibronectin type III domain and a 75 aa Ig C2-like region. The expression of p40 is induced by substances such as LPS and CpG that activate antigen-presenting cells. Besides being found as a component of IL-12 or IL-23, free p40 monomers and homodimers are also secreted by cells expressing p40. Feline p40 shares 94%, 85%, 84%, 65%, and 65% aa sequence identity with canine, human, porcine, rat and mouse p40, respectively. Cells known to express p40 include macrophages, dendritic cells, monocytes, Langerhans cells, neutrophils, keratinocytes, plasmacytoid dendritic cells, and microglia. From cells that express both the p35 and p40 subunits (dendritic cells, monocytes, and CHO cells), the amount of free p40 secreted is 10‑1000 fold more than the heterodimeric IL-12. The high-affinity IL-12 receptor complex that transduces IL-12 signals is composed of a 100 kDa ligand-binding subunit (IL-12 R beta 1) and a 130 kDa signal transducing subunit (IL-12 R beta 2). Similarly, the high-affinity IL-23 signaling receptor complex is composed of the shared IL-12 R beta 1 and the unique IL-23 R, a novel gp130-like protein. Both the monomeric and the dimeric free p40 can bind to the IL-12 R beta 1 and function as antagonists of IL-12 or IL-23. However, the monomeric p40 binds IL-12 R beta 1 with lower affinity and is less potent as an IL-12 antagonist. Homodimeric mouse p40 has also been shown to have agonistic functions similar to IL-12, inducing nitric oxide expression and NF kappa B activation in mouse primary microglia and peritoneal macrophages. The molecular mechanism for the agonistic effects of homodimeric p40 has not been determined (1‑6).

  • References:
    1. Buttner, M. et al. (1997) Cytokine 10:241.
    2. Park, A.Y. and P. Scott (2001) Scand. J. Immunol. 53:529. 
    3. Trinchieri, G. et al. (2003) Immunity 19:641. 
    4. Brombacher, F. et al. (2003) Trends Immunol. 24:207.
    5. Lankford, C.S. and D.M. Frucht, 2003, J. Leukoc. Biol. 73:49.
    6. Abdi, K. (2002) Scand. J. Immunol. 5:1.
  • Long Name:
    Interleukin 12/Interleukin 23 p40
  • Entrez Gene IDs:
    3593 (Human); 16160 (Mouse); 64546 (Rat); 403976 (Canine); 493741 (Feline)
  • Alternate Names:
    CLMF2; IL12 p40; IL-12 p40; IL12B; IL-12B; NKSF2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Efficient plant-based production of chicken interleukin-12 yields a strong immunostimulatory cytokine.
    Authors: Medrano G, Dolan MC, Stephens NT, McMickle A, Erf G, Radin D, Cramer CL
    J. Interferon Cytokine Res., 2010;30(3):143-54.
    Species: Chicken
    Sample Type: Recombinant Protein
    Application: WB
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