Canine IL-6 Antibody

Catalog # Availability Size / Price Qty
AF1609
AF1609-SP
Cell Proliferation Induced by IL‑6 and Neutralization by Canine IL‑6 Antibody.
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Citations (2)
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Canine IL-6 Antibody Summary

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 either lyophilized or 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

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
0.2-0.8 µg/mL 

Use in combination with:

Detection Reagent: Canine IL‑6 Biotinylated Antibody (Catalog # BAF1609)

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.

Scientific Data

Neutralization Cell Proliferation Induced by IL‑6 and Neutralization by Canine IL‑6 Antibody. View Larger

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 antibody in CLL-1390 Canine Cell Line antibody by Flow Cytometry. View Larger

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 antibody in Canine PBMCs by Immunocytochemistry (ICC). View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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); 102138971 (Cynomolgus Monkey); 100034196 (Equine); 493687 (Feline); 463288 (Primate); 100008733 (Rabbit)
Alternate Names
B-cell differentiation factor; B-cell stimulatory factor 2; BSF2; BSF-2; CDF; CTL differentiation factor ; HSF; hybridoma growth factor; IFNB2; IFN-beta-2; IL6; IL-6; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A

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Citations for Canine IL-6 Antibody

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.

2 Citations: Showing 1 - 2
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  1. Initial Pulmonary Respiration Causes Massive Diaphragm Damage and Hyper-CKemia in Duchenne Muscular Dystrophy Dog
    Authors: Akinori Nakamura, Masanori Kobayashi, Mutsuki Kuraoka, Katsutoshi Yuasa, Naoko Yugeta, Takashi Okada et al.
    Scientific Reports
  2. Therapeutic intervention using a Smad7-based Tat protein to treat radiation- induced oral mucositis
    Authors: Boss MK, Ke Y, Bian L Et al.
    International journal of radiation oncology, biology, physics

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