Immunoassays | Inflammation & Cytokines

Erenna® IL-4 Immunoassay Kit (Cat# 03-0052-xx)

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Introduction

The Erenna® IL-4 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IL-4 in human serum and plasma samples. A capture antibody specific for human IL-4 has been pre-coated onto paramagnetic microparticles (MPs). The user pipettes MPs, standards, and samples into uncoated microplate wells. During incubation, the IL-4 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-4 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-4 present in the sample when captured. The amount of IL-4 in unknown samples is interpolated from a standard curve.

Assay Performance


TABLE 1. Analytical sensitivity of the Erenna® IL-4 Immunoassay Kit1
Lower Limit of Detection 0.003 pg/mL
Lower Limit of Quantification2 0.02 pg/mL
Upper Limit of Quantification 10 pg/mL
Low-end CV% Range 2 – 11%
Low-end CV% Average 6%
Recommended Sample Volume 100
Minimum Sample Volume Required3 100

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-4] in a healthy reference


FIGURE 1. [IL-4] in EDTA plasma from 10 healthy donors (6 samples < LLoQ), with median and interquartile range. The Erenna® IL-4 Immunoassay Kit can quantify IL-4 in healthy subjects, who have a median [IL-4] of 0.02pg/mL that is above the detection limit of 0.003 pg/mL.

FIGURE 2. Erenna® IL-4 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-4 (IL-4), is a cytokine that is a key regulator in humoral and adaptive immunity. IL-4 induces differentiation of naive helper T-cells (Th0 cells) to Th2 cells. It has many biological roles, including the stimulation of activated B-cell and T-cell proliferation, and the differentiation of CD4+ T-cells into Th2 cells. Further, IL-4 controls the production of IgE, expands IL-4 producing T-cell subsets, and stabilizes effector cell functions. IL-4 has great therapeutic potential due to its role in the development of allergic inflammatory responses. IL-4 also has shown to have promise in drug targeting for cancer.


TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-4
UniProtKB/Swiss-Prot ID: P05112
Protein-protein interaction database: P05112
Alternative Names: B-cell stimulatory factor, Binetrakin Lymphocyte stimulatory factor 1
Gene Names: IL4
Synonym Gene Names:
Function: Participates in at least several B-cell activation processes as well as of other cell types. It is a costimulator of DNA-synthesis. It induces the expression of class II MHC molecules on resting B-cells. It enhances both secretion and cell surface expression of IgE and IgG1. It also regulates the expression of the low affinity Fc receptor for IgE (CD23) on both lymphocytes and monocytes.
Biological Processes: B-cell activation
Molecular Function: Cytokine, Growth factor

* Source Info from www.uniprot.org.


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