Human IL-6 Antibody

R&D Systems | Catalog # AF-206-NA

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), Affinity Assay, Bioassay, ELISA Capture, ELISA Detection, ELISA Development, Proximity Ligation Assay, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-6 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human IL-6
Pro29-Met212
Accession # P05231

Specificity

Detects human IL-6 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse 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 Human IL-6 Antibody

IL-6 antibody in Human PBMCs by Immunocytochemistry (ICC).

IL‑6 in Human PBMCs.

IL-6 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with LPS and monensin using Goat Anti-Human IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-206-NA) at 10 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 counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by IL‑6 and Neutralization by Human IL‑6 Antibody.

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

Recombinant Human IL-6 (Catalog # 206-IL) stimulates proliferation in the T1165.85.2.1 mouse plasmacytoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-6 (2.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-206-NA). The ND50 is typically ≤ 125 ng/mL.
Detection of Human IL-6 by Immunohistochemistry

Detection of Human IL-6 by Immunohistochemistry

Breast DCIS cells overexpress proinflammatory markers. Representative images of immunohistochemistry targeting IL-6 protein in normal breast tissue (a), and breast DCIS (b) (N = 61). c and d Hematoxylin staining of serial sections from tissue shown in panel A and B. All images are 20X magnification. Scale bar equals 100 μm. e Evaluation of IL-6 gene expression in DCIS cells via qRT-PCR; the isogenic MCF10.DCIS cells and the non-isogenic SUM102 cell line were analyzed against the non-transformed MCF10A cell line (N = 3). f Secretion of IL-6 protein from DCIS cell lines and non-transformed MCF10A cells as determined by ELISA. *P < 0.05, Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Block/Neutralize

Detection of Human IL-6 by Block/Neutralize

IL-6 neutralizing antibody (nAb) inhibits growth of MCF10.DCIS structures. Representative contiguous 16-tiled DIC images of MCF10.DCIS cells grown in MAME cultures for 8 days. MAME culture of MCF10.DCIS cells treated with an isotype-matched antibody against IgG (control) (8 days) (a), or 1 μg /ml IL-6 nAb (b) (8 days). MCF10.DCIS cells treated with IL-6 nAb followed by a 48 h treatment-free recovery period prior to imaging on day 10 (c). N = 3, Scale bars, 200 μm. d Diameter of MCF10.DCIS structures in the presence of control antibody, IL-6 nAb, or 48 h recovery from IL-6 nAb. N = 20-40 measurements /tiled DIC image (N = 3). ****P ≤ 0.0001, **P ≤ 0.01. e Evaluation of gene expression of invasive tumor cell markers in MAME cultures treated with IL-6 nAb (N = 3). Fold difference as compared to control cultures. Dashed line indicates 2-fold threshold. Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Block/Neutralize

Detection of Human IL-6 by Block/Neutralize

IL-6 nAb inhibits CAF-stimulated MCF10.DCIS structure growth and results in altered morphology. Contiguous 16-tile DIC image of CAFs: WS12Ti (A, B) or CAF40TKi (c, d) co-cultured with MCF10.DCIS cells for 8 days in the presence of isotype matched anti-IgG (a, c) or 1 μg /ml IL-6 nAb (b, d). e Perimeter measurements from DCIS structures in MAME culture in the presence of anti-IgG or IL-6 nAb (N = 100-140 measurements from 6 contiguous tiled DIC images (N = 3). Fluorescent tiled images of CAF40TKi fibroblasts (unlabeled) cultured with MCF10.DCIS-RFP cells in the presence of anti-IgG (f) or 1 μg /ml IL-6 nAb (g). h Quantification of total volume of DCIS-RFP structures in co-culture with CAF40TKi fibroblasts (ratio-paired t-test, N = 3). ***P ≤ 0.001, **P ≤ 0.01; Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Block/Neutralize

Detection of Human IL-6 by Block/Neutralize

IL-6 neutralizing antibody (nAb) inhibits growth of MCF10.DCIS structures. Representative contiguous 16-tiled DIC images of MCF10.DCIS cells grown in MAME cultures for 8 days. MAME culture of MCF10.DCIS cells treated with an isotype-matched antibody against IgG (control) (8 days) (a), or 1 μg /ml IL-6 nAb (b) (8 days). MCF10.DCIS cells treated with IL-6 nAb followed by a 48 h treatment-free recovery period prior to imaging on day 10 (c). N = 3, Scale bars, 200 μm. d Diameter of MCF10.DCIS structures in the presence of control antibody, IL-6 nAb, or 48 h recovery from IL-6 nAb. N = 20-40 measurements /tiled DIC image (N = 3). ****P ≤ 0.0001, **P ≤ 0.01. e Evaluation of gene expression of invasive tumor cell markers in MAME cultures treated with IL-6 nAb (N = 3). Fold difference as compared to control cultures. Dashed line indicates 2-fold threshold. Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Block/Neutralize

Detection of Human IL-6 by Block/Neutralize

IL-6 nAb inhibits CAF-stimulated MCF10.DCIS structure growth and results in altered morphology. Contiguous 16-tile DIC image of CAFs: WS12Ti (A, B) or CAF40TKi (c, d) co-cultured with MCF10.DCIS cells for 8 days in the presence of isotype matched anti-IgG (a, c) or 1 μg /ml IL-6 nAb (b, d). e Perimeter measurements from DCIS structures in MAME culture in the presence of anti-IgG or IL-6 nAb (N = 100-140 measurements from 6 contiguous tiled DIC images (N = 3). Fluorescent tiled images of CAF40TKi fibroblasts (unlabeled) cultured with MCF10.DCIS-RFP cells in the presence of anti-IgG (f) or 1 μg /ml IL-6 nAb (g). h Quantification of total volume of DCIS-RFP structures in co-culture with CAF40TKi fibroblasts (ratio-paired t-test, N = 3). ***P ≤ 0.001, **P ≤ 0.01; Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Immunohistochemistry

Detection of Human IL-6 by Immunohistochemistry

Breast DCIS cells overexpress proinflammatory markers. Representative images of immunohistochemistry targeting IL-6 protein in normal breast tissue (a), and breast DCIS (b) (N = 61). c and d Hematoxylin staining of serial sections from tissue shown in panel A and B. All images are 20X magnification. Scale bar equals 100 μm. e Evaluation of IL-6 gene expression in DCIS cells via qRT-PCR; the isogenic MCF10.DCIS cells and the non-isogenic SUM102 cell line were analyzed against the non-transformed MCF10A cell line (N = 3). f Secretion of IL-6 protein from DCIS cell lines and non-transformed MCF10A cells as determined by ELISA. *P < 0.05, Student’s t-test; mean ± SD Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26268945), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-6 by Western Blot

Detection of Human IL-6 by Western Blot

IL-1 alpha upregulates miR-146a in senescent cells.(A) Northern analysis for miR-146a levels in damage-induced senescent HCA2 cells treated with neutralizing antibodies against IL1-alpha and IL1-beta. HCA2 cells (PD 35) were used and induced to senesce by treatment with bleomycin. Cells were harvested for RNA 11 days later. Details of the procedure are described in ‘Experimental Procedures. (B) Western analysis for IL-6 in damage-induced senescent HCA2 cells treated with neutralizing antibodies to IL-1 alpha and IL-1 beta. CM were harvested 11 days after bleomycin treatment. \ Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20148189), licensed under a CC-BY license. Not internally tested by R&D Systems.
Human IL-6 ELISA Standard Curve

Human IL-6 ELISA Standard Curve

Recombinant Human IL‑6 (Catalog # 206-IL) was serially diluted and captured by Mouse Anti-Human/Primate IL‑6 Monoclonal Antibody (Catalog # MAB206) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-206-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human IL-6 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells treated with LPS and monensin

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 ≤ 125 ng/mL in the presence of 2.5 ng/mL Recombinant Human IL‑6.

Reviewed Applications

Read 6 reviews rated 4.5 using AF-206-NA in the following applications:

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.

Calculators

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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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 human IL-6 is 183 amino acids (aa) in length expressed as a 22-28 kDA molecular weight protein. IL-6 shares 39% aa sequence identity with mouse and rat IL-6. 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

P05231

Additional IL-6 Products

Product Documents for Human IL-6 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IL-6 Antibody

For research use only

Citations for Human IL-6 Antibody

Customer Reviews for Human IL-6 Antibody (6)

4.5 out of 5
6 Customer Ratings
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Customer Images

Showing  1 - 5 的 6 reviews Showing All
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  • Human IL-6 Antibody
    Name: Zoia Levashova
    Application: MSD Assay
    Sample Tested: Vitreous humor
    Species: Cynomolgus Monkey
    Verified Customer | Posted 11/01/2018
    After labeling with Sulfo-Tag, used as a detection reagent in MSD assay (Meso Scale Diagnostics LLC). Paired with biotinylated IL-6 antibody (MAB206) as a capture reagent. A standard curve with recombinant human IL-6 from (Cat# 206-IL/CF) is shown (1.6-25,000 pg/ml).
    Human IL-6 Antibody AF-206-NA
  • Human IL-6 Antibody
    Name: Anonymous
    Application: Cell depletion
    Sample Tested: Pancreatic cancer cells
    Species: Human
    Verified Customer | Posted 05/11/2018
  • Human IL-6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Whole cell lysates
    Species: Human
    Verified Customer | Posted 12/22/2016
    Endothelial cells treated with TNF-a (50 ng/mL) for 1 hour.
    Human IL-6 Antibody AF-206-NA
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 21784845
    Species: Human
    Verified Customer | Posted 01/06/2015
  • Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: HEK293 cells transfected with hIL6
    Species: Human
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: hIL6 transfected cells
    Species: Human
    Verified Customer | Posted 12/19/2014

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FAQs for Human IL-6 Antibody

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  • Q: What band size does AF-206-NA detect in Western blot?

    A: AF-206-NA detects a 20-22 kDa band in lysate in Western blot.

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