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Immunoassays | Inflammation & Cytokines

Erenna® IL-10 Immunoassay Kit (Cat# 03-0056-xx)

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The Erenna® IL-10 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IL-10 in human plasma samples. A capture antibody specific for human IL-10 has been pre-coated onto paramagnetic microparticles (MPs). The user pipettes MPs, standards, and samples into uncoated microplate wells. During incubation, the IL-10 present in the sample binds to the capture antibody on the coated MPs. 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-10 that has been captured onto the MPs. 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-10 present in the sample when captured. The amount of IL-10 in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-10 Immunoassay Kit1
Lower Limit of Detection 0.06 pg/mL
Lower Limit of Quantification2 0.39 pg/mL
Upper Limit of Quantification 400 pg/mL
Low-end CV% Range 1 - 12%
Low-end CV% Average 6%
Recommended Sample Volume 100 μL
Minimum Sample Volume Required3 39 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-10] in a healthy reference

FIGURE 1. [IL-10] in EDTA plasma from 12 healthy donors, with median and interquartile range. The Erenna® IL-10 Immunoassay Kit reliably quantifies IL-10 in healthy subjects, who have a median [IL-10] of 16.11 pg/mL that is well above the detection limit of 0.48 pg/mL.

FIGURE 2. Erenna® IL-10 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-10 (IL-10) is a cytokine with both stimulatory and suppressive effects on the immune system. The protein originates from several cell lines, including Th2 cells, mast cells, macrophages, monocytes and B-cells. Its main functions are to impede inflammatory responses and to inhibit synthesis of other pro-inflammatory cytokines, such as IL-2, IL-3, GM-CSF, IFN-γ and TNF-α. Besides its anti-inflammatory capabilities, IL-10 also regulates the production of natural killer cells and lymphocytes. Due to its multiple roles, proper function of IL-10 is required to maintain a healthy immune system. Improper function of IL-10 can result in disease development, as over expression of IL-10 has been linked to lymphoma, whereas IL-10 deficiency has implications for Crohn’s disease.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-10
UniProtKB/Swiss-Prot ID: P22301
Protein-protein interaction database: P22301
Alternative Names: Cytokine synthesis inhibitory factor
Gene Names: IL10
Synonym Gene Names:
Function: Inhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells.
Biological Processes: B cell differentiation, B cell proliferation, T-helper 2 type immune response, anti-apoptosis, cell-cell signaling, cytoplasmic sequestering of NF-kappaB, inflammatory response, leukocyte chemotaxis, negative regulation of B cell proliferation, negative regulation of MHC class II biosynthetic process, negative regulation of T cell proliferation
Molecular Function: cytokine activity, growth factor activity, interleukin-10 receptor binding

* Source Info from www.uniprot.org.

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