Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Neutralization, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, Cell Culture, Immunoassay Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1936
View all IL-6 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human IL‑6
Val30-Met212
Accession # P05231

Specificity

Detects human IL-6 in direct ELISAs. Does not cross-react with recombinant IL-6 from mouse, rat, or pig.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.20 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human IL‑6 Antibody

IL-6 antibody in Human Appendix by Immunohistochemistry (IHC-P).

IL‑6 in Human Appendix.

IL-6 was detected in immersion fixed paraffin-embedded sections of human appendix using Mouse Anti-Human IL-6 Monoclonal Antibody (Catalog # MAB2061) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of IL-6 antibody in Human PBMCs antibody by Flow Cytometry.

Detection of IL‑6 in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) treated with 100 ng/mL LPS for 24 hours were stained with Mouse Anti-Human IL-6 Monoclonal Antibody (Catalog # MAB2061, filled histogram) or isotype control antibody (MAB004, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). View our protocol for Staining Intracellular Molecules.
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 (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 Mouse Anti-Human IL-6 Monoclonal Antibody (Catalog # MAB2061). The ND50 is typically 8.00-80.0 ng/mL.

Detection of IL‑6 in Human PBMCs.

IL‑6 was detected in immersion fixed Human PBMCs stimulated with LPS (positive control) and absent in untreated human PBMCs (negative control) using Mouse Anti-Human IL‑6 Monoclonal Antibody (Catalog # MAB2061) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of IL‑6 in PBMC monocytes by Flow Cytometry.

PBMCs stimulated with 100 ng/ml LPS and Brefeldin A for 24 hrs (A) or unstimulated (B) were stained with Mouse Anti-Human IL‑6 Monoclonal Antibody (Catalog # MAB2061) and Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (1X) (Catalog # FC004) and permeabilized with Saponin. View our protocol for Staining Intracellular Molecules.
Detection of IL-6 by Immunocytochemistry/ Immunofluorescence

Detection of IL-6 by Immunocytochemistry/ Immunofluorescence

Effects of topo I-reactive B cells on the differentiation of CD4+ T cells in systemic sclerosis (SSc) patients.B cells with low affinity for topo I and those with high affinity for topo I as well as topo I-non-reactive B cells were obtained from anti-topo I antibody-positive SSc patients (n = 111). These B cells were co-cultured with CD4+ T cells. After 48 hr of co-culture, mRNA was extracted from these cells and FoxP3 and ROR gamma t expression levels were examined by real-time RT-PCR. The results were presented when 96-well plates were used as a co-culture site (A) and when microculture plates (B) were used (C). These cells were further co-cultured on microculture plates, and the protein expression of CD4, CD20, FoxP3, and ROR gamma t was confirmed by fluorescent cell staining and signal intensity was determined by ImageJ (D). Similarly, co-culture in microculture plates in the presence of anti-IL-10 (10 μg/ml), anti-IL-35 (5 μg/ml), anti-IL-6 (1 μg/ml), or anti-IL-23 (5 μg/ml) antibodies (Abs) was conducted, followed by fluorescent cell staining (E). These results represented seven experiments. The bar graphs show the mean + SD. Original magnification, ×1000. *p<0.05.Figure 3—source data 1.Source file for the effects of topo I-reactive B cells on the differentiation of CD4+ T cells in systemic sclerosis (SSc) patients.This archive contains all data used for the quantitative analysis shown in Figure 3.Source file for the effects of topo I-reactive B cells on the differentiation of CD4+ T cells in systemic sclerosis (SSc) patients.This archive contains all data used for the quantitative analysis shown in Figure 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34854378), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human IL‑6 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

3-15 µg/mL
Sample: Immersion fixed Human PBMCs stimulated with LPS (positive control) and untreated human PBMCs (negative control)

Immunohistochemistry

1-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human appendix

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: PBMCs treated +/- LPS (see details below) 

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 8.00-80.0 ng/mL in the presence of 2.5 ng/mL Recombinant Human IL‑6.

Reviewed Applications

Read 2 reviews rated 4.5 using MAB2061 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS and NaCl with Trehalose. See Certificate of Analysis for details.
*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

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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 (2)

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  • Human IL-6 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Colon tissue
    Species: Human
    Verified Customer | Posted 06/18/2022
    Human IL‑6 Antibody MAB2061
  • Human IL‑6 Antibody
    Name: Jason Thall
    Application: ELISA
    Sample Tested: Mouse ear fibroblast
    Species: Human
    Verified Customer | Posted 04/29/2016

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