Canine IL-6 Antibody

  
  • Species Reactivity
    Canine
  • Specificity
    Detects canine IL-6 in ELISAs and Western blots. In sandwich ELISAs, less than 5% cross-reactivity with recombinant feline IL‑6 and recombinant porcine IL‑6 is observed, and less than 0.2% cross-reactivity with recombinant human IL-6, recombinant mouse IL‑6, recombinant rat IL‑6, recombinant cotton rat IL‑6, and recombinant equine IL‑6 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine IL‑6
    Thr23-Met207
    Accession # P41323
  • 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 Canine IL‑6 (Catalog # 1609-CL)
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
    • Canine IL-6 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Canine IL‑6 Antibody (Catalog # AF1609)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Canine IL‑6 Biotinylated Antibody (Catalog # BAF1609)
  • ELISA Standard
     
    Recombinant Canine IL-6 Protein (Catalog # 1609-CL)
  • Neutralization
    Measured by its ability to neutralize IL‑6-induced proliferation in the T1165.85.2.1 mouse plasmacytoma cell line. The Neutralization Dose (ND50) is typically 0.4-2.0 µg/mL in the presence of 50 ng/mL Recombinant Canine IL‑6.
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‑6 and Neutralization by Canine IL‑6 Antibody.

Recombinant Canine IL‑6 (Catalog # 1609-CL) stimulates proliferation in the T1165.85.2.1 mouse plasmacytoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Canine IL‑6 (50 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine IL‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1609). The ND50 is typically 0.4-2.0 µg/mL.

Intracellular Staining by Flow Cytometry
Detection of IL‑6 in CLL‑1390 Canine Cell Line by Flow Cytometry.

CLL‑1390 canine leukocytic round cell neoplasia cell line was stained with Goat Anti-Canine IL‑6 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1609, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Immunocytochemistry
IL‑6 in Canine PBMCs. IL‑6 was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) stimulated with PHA using Goat Anti-Canine IL‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1609) 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-6

Interleukin 6 (IL-6) is a pleiotropic alpha -helical cytokine that plays important roles in acute phase reactions, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6 activity is central to the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. It is secreted by multiple cell types as a 22-28 kDa phosphorylated and variably glycosylated molecule (1-4). Mature canine IL-6 is 187 amino acids (aa) in length and shares 76%, 59%, 38%, and 40% aa sequence identity with feline, human, mouse, and rat IL-6, respectively (5). IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R, triggering IL-6 R association with gp130 and gp130 dimerization (6). gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM (7). Soluble forms of IL-6 R are generated by both alternate splicing and proteolytic cleavage (3). In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R elicit responses from gp130-expressing cells that lack cell surface IL-6 R (3). Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous while that of IL-6 R is predominantly restricted to hepatocytes, leukocytes, and lymphocytes (3). Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R but not from other cytokines that utilize gp130 as a coreceptor (4, 8).

  • References:
    1. Van Snick, J. (1990) Annu. Rev. Immunol. 8:253.
    2. Hodge, D.R. et al. (2005) Eur. J. Cancer 41:2502.
    3. Jones, S.A. (2005) J. Immunol. 175:3468.
    4. Rose-John, S. et al. (2006) J. Leukoc. Biol. 80:227.
    5. Kukielka, G.L. et al. (1995) Circulation 92:1866.
    6. Murakami, M. et al. (1993) Science 260:1808.
    7. Muller-Newen, G. (2003) Sci. STKE 2003:PE40.
    8. Mitsuyama, K. et al. (2006) Clin. Exp. Immunol. 143:125.
  • Long Name:
    Interleukin 6
  • Entrez Gene IDs:
    3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 100034196 (Equine); 493687 (Feline); 100008733 (Rabbit)
  • Alternate Names:
    B cell stimulatory factor-2; B-cell differentiation factor; BSF-2; BSF2CTL differentiation factor; CDF; HGFHSFIFNB2Hybridoma growth factor; IFN-beta-2; IL6; IL-6; IL-6B-cell stimulatory factor 2; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A
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