详细说明
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 # P41323Formulation
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
ConcentrationSample
Western Blot
0.1 µg/mL
Recombinant Canine IL‑6 (Catalog # )
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/10 6 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 # )
ELISA Detection (Matched Antibody Pair)
0.1-0.4 µg/mL
Canine IL‑6 Biotinylated Antibody (Catalog # )
ELISA Standard
Recombinant Canine IL-6 Protein (Catalog # )
Neutralization
Measured by its ability to neutralize IL‑6-induced proliferation in the T1165.85.2.1 mouse plasmacytoma cell line. The Neutralization Dose (ND 50) 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. are available in the Technical Information section on our website.
Data Examples
Neutralization | Cell Proliferation Induced by IL‑6 and Neutralization by Canine IL‑6 Antibody. Recombinant Canine IL‑6 (Catalog # ) 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 # , open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # ). 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 # ) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for . |
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:
Van Snick, J. (1990) Annu. Rev. Immunol. 8:253.
Hodge, D.R. et al. (2005) Eur. J. Cancer 41:2502.
Jones, S.A. (2005) J. Immunol. 175:3468.
Rose-John, S. et al. (2006) J. Leukoc. Biol. 80:227.
Kukielka, G.L. et al. (1995) Circulation 92:1866.
Murakami, M. et al. (1993) Science 260:1808.
Muller-Newen, G. (2003) Sci. STKE 2003:PE40.
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