Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry, Bioassay, In vivo assay, Fuorescense Microscopy, Immunodepletion

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all IL-6 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat IL-6
Phe25-Thr211
Accession # P20607

Specificity

Detects rat and mouse IL-6 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant cotton rat IL‑6 is observed, and less than 1% cross-reactivity with recombinant human IL-6 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Rat IL-6 Antibody

Detection of Rat IL‑6 by Western Blot.

Western blot shows lysates of NR8383 rat alveolar macrophage cell line untreated (-) or treated (+) with LPS. PVDF membrane was probed with 1 µg/mL of Goat Anti-Rat IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF506) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-6 at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL-6 in NR8383 Rat Cell Line.

IL-6 was detected in immersion fixed NR8383 rat alveolar macrophage cell line using Goat Anti-Rat IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF506) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cyoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Cell Proliferation Induced by IL‑6 and Neutralization by Rat IL‑6 Antibody.

Recombinant Rat IL-6 (Catalog # 506-RL) stimulates proliferation in the T1165.85.2.1 mouse plasmacytoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL-6 (0.6 ng/mL) is neutralized (green line) by increasing concentrations of Rat IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF506). The ND₅₀ is typically

Detection of Porcine IL-6 by Immunohistochemistry-Frozen

Diverse localization of measured cytokines in P7 brains. Representative pictures (100× magnification) of immunohistochemical stainings of IL-1 beta (Α), IL-10 (B), IL-6 (C) and TNF-alpha (D). Cytokines are stained with DAB (brown color) and counterstained with cresyl violet. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23351591), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rat IL-6 by Immunocytochemistry/ Immunofluorescence

Rat IL-6 ELISA Standard Curve

Recombinant Rat IL-6 (Catalog # 506-RL) was serially diluted and captured by Mouse Anti-Rat IL-6 Monoclonal Antibody (Catalog # MAB506) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF506) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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Applications for Rat IL-6 Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed NR8383 rat alveolar macrophage cell line

Western Blot

1 µg/mL
Sample: NR8383 rat alveolar macrophage cell line treated with LPS

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 <0.5 µg/mL in the presence of 0.6 ng/mL Recombinant Rat IL‑6.

Reviewed Applications

Read 1 review rated 4 using AF506 in the following applications:

Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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, phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. Mature rat IL-6 is 187 amino acids (aa) in length that is typically expressed as a 22-28 kDA molecular weight protein. Rat IL-6 shares 39% and 85% aa sequence identity with human and mouse IL-6, respectively. Alternative splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties. IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R alpha) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R alpha, triggering IL-6 R alpha association with gp130 and gp130 dimerization. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha. 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 alpha is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R alpha but not from other cytokines that use gp130 as a co-receptor. IL-6, along with TNF-alpha and IL-1, function to drive the acute inflammatory response and the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 can also function as an anti-inflammatory molecule, as in skeletal muscle where it is secreted in response to exercise. In addition, it enhances hematopoietic stem cell proliferation and the differentiation of Th17 cells, memory B cells, and plasma cells.

Long Name

Interleukin 6

Alternate Names

BSF-2, BSF2, IFNB2, IL6, MGI-2A

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)

Gene Symbol

IL6

UniProt

P20607

Additional IL-6 Products

Product Documents for Rat IL-6 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Rat IL-6 Antibody

For research use only

Citations
Reviews

Rat IL-6 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF506

Key Product Details

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Species Reactivity

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Cited:

Applications