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Immunoassays | Inflammation & Cytokines

Erenna® IL-13 Immunoassay Kit (Cat# 03-0069-xx)

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The Erenna® IL-13 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IL-13 in human serum and plasma samples. A capture antibody specific for human IL-13 has been pre-coated onto paramagnetic microparticles (MPs). The user pipettes MPs, standards, and samples into uncoated microplate wells. During incubation, the IL-13 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-13 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-13 present in the sample when captured. The amount of IL-13 in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-13 Immunoassay Kit1
Lower Limit of Detection 0.01 pg/mL
Lower Limit of Quantification2 0.39 pg/mL
Upper Limit of Quantification 100 pg/mL
Low-end CV% Range 1 - 7%
Low-end CV% Average 5%
Recommended Sample Volume 100 μL
Minimum Sample Volume Required3 50 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-13] in a healthy reference

FIGURE 1. [IL-13] in EDTA plasma from 10 healthy donors, with median and interquartile range.

FIGURE 2. The Erenna® IL-13 Immunoassay Kit low-end standard 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-13 (IL-13) is a cytokine that is crucial for mediating allergic responses. Predominately expressed by Th2 cells, IL-13 is also secreted by cell types such as mast cells, NK cells, and dentritic cells. Both IL-13 and IL-4 are found at elevated levels in the asthmatic lung and are thought to be key regulators of airway inflammation. IL-13 functions similarly to IL-4 in that both can signal through the heterodimer receptor IL-4Rα - IL-13Rα1 and activate STAT-6 downstream. However, studies have suggested that IL-13 operates independently of IL-4 with regard to asthma regulation. Thus, IL-13 has become a promising therapeutic target for allergic diseases. Several anti-IL-13 antibodies are being evaluated as treatment for bronchial asthma.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-13
UniProtKB/Swiss-Prot ID: P35225
Protein-protein interaction database:
Alternative Names:
Gene Names: IL13
Synonym Gene Names: NC30
Function: Cytokine. Inhibits inflammatory cytokine production. Synergizes with IL2 in regulating interferon-gamma synthesis. May be critical in regulating inflammatory and immune responses.
Biological Processes: extracellular space Inferred from sequence or structural similarity. soluble fraction
Molecular Function: cytokine activity.

* Source Info from www.uniprot.org.

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