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Immunoassays | Inflammation & Cytokines

Erenna® IL-6 Immunoassay Kit (Cat# 03-0010-xx)

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The Erenna® IL-6 Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure IL-6 in human plasma and serum samples. A capture antibody specific for human IL-6 has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards, and samples into uncoated microplate wells. During incubation, the IL-6 present in the sample binds to the capture antibody on the coated MP. Unbound molecules are washed away during the subsequent buffer exchange and wash steps. Fluor-labeled detection antibody is added to each well and incubated. This detection antibody recognizes and binds to IL-6 that has been captured onto the MP. During the following wash step the MPs are transferred to a clean plate. Elution buffer is then added and incubated. The elution buffer dissociates the bound protein sandwich from the MP surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final microplate. The plate is loaded into the Erenna System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IL-6 present in the sample when captured. The amount of IL-6 in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-6 Immunoassay Kit1
Lower Limit of Detection 0.005 pg/mL
Lower Limit of Quantification2 0.01 pg/mL
Upper Limit of Quantification 50 pg/mL
Low-end CV% Range 2 - 8%
Low-end CV% Average 5%
Recommended Sample Volume 75 μL
Minimum Sample Volume Required3 1 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-6] in a healthy reference

FIGURE 1. [IL-6] in EDTA plasma from 20 healthy donors, with median and interquartile range. The Erenna® IL-6 Immunoassay Kit reliably quantifies IL-6 in healthy subjects, who have a median [IL-6] of 1.2 pg/mL that is well above the detection limit of 0.005 pg/mL.

FIGURE 2. Erenna® IL-6 Immunoassay Kit low-end standard curve signal (left) and curve fit (right).

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Interleukin-6 (IL-6) is a pro-inflammatory cytokine secreted by T cells and macrophages to stimulate immune response to trauma, especially burns or other tissue damage leading to inflammation. IL-6 is also secreted by macrophages in response to specific microbial molecules, referred to as pathogen associated molecular patterns (PAMPs), which trigger the innate immune response and initiate inflammatory cytokine production. IL-6 is one of the most important mediators of fever and of the acute phase response. IL-6 is also called a “myokine,” a cytokine produced from muscle that increases in response to muscle contraction. Additionally, steoblasts secrete IL-6 to stimulate osteoclast formation.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-6
UniProtKB/Swiss-Prot ID: P05231
Protein-protein interaction database: P05231
Alternative Names: B-cell stimulatory factor 2, CTL differentiation factor, Hybridoma growth factor, Interferon beta-2
Gene Names: IL6
Synonym Gene Names: IFNB2
Function: Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance.
Biological Processes: acute-phase response, defense response to Gram-negative bacterium, defense response to Gram-positive bacterium, defense response to virus, endocrine pancreas development, glucagon secretion, hepatic immune response, humoral immune response, interleukin-6-mediated signaling pathway, monocyte chemotaxis, negative regulation of apoptosis, negative regulation of cell proliferation, negative regulation of chemokine biosynthetic process, negative regulation of collagen biosynthetic process, negative regulation of fat cell differentiation, negative regulation of lipid storage, neuron projection development, neutrophil apoptosis, neutrophil mediated immunity, platelet activation, positive regulation of B cell activation, positive regulation of MAPKKK cascade, positive regulation of T cell proliferation, positive regulation of acute inflammatory response,
Molecular Function: cytokine activity, growth factor activity, interleukin-6 receptor binding

* Source Info from www.uniprot.org.

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