Feline IL-6 Antibody

Catalog #: AF2305 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF2305
AF2305-SP

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Feline IL-6 Antibody Summary

Species Reactivity

Feline

Specificity

Detects feline IL‑6 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant canine IL‑6, recombinant porcine IL‑6 and recombinant human IL‑6 is observed and less than 10% cross-reactivity with recombinant mouse
IL‑6 and recombinant rat IL‑6 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant feline IL‑6
Thr28-Met208 (Glu133Lys)
Accession # P41683

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

<0.20 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‑6 (Catalog # 2305-FL)

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize IL‑6-induced proliferation in the T1165.85.2.1 mouse plasmacytoma cell line. Nordan, R. P. and M. Potter (1986) Science 233:566. The Neutralization Dose (ND₅₀) is typically 0.2-0.6 µg/mL in the presence of 10 ng/mL Recombinant Feline 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

Immunocytochemistry

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IL‑6 in Feline PBMCs. IL-6 was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-Feline IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2305) 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.

Neutralization

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Cell Proliferation Induced by IL‑6 and Neutralization by Feline IL‑6 Antibody. Recombinant Feline IL-6 (Catalog # 2305-FL) stimulates proliferation in the T1165.85.2.1 mouse plasmacytoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Feline IL-6 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Feline IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2305). The ND₅₀ is typically 0.2-0.6 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 kDa‑28 kDa phosphorylated and variably glycosylated molecule (1‑4). Mature feline IL-6 is 181 amino acids (aa) in length and shares 76%, 58%, 39%, and 41% aa sequence identity with canine, 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.
Ohashi, T. et al. (1993) J. Vet. Med. Sci. 5:941.
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); 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