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Immunoassays | Inflammation & Cytokines

Erenna® IL-1α Immunoassay Kit (Cat# 03-0050-xx)

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Introduction

The Erenna® IL-1α Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure IL-1α in plasma and serum samples. A capture antibody specific for IL-1α has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards, and samples into uncoated microplate wells. During incubation, the IL-1α 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-1α 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-1α present in the sample when captured. The amount of IL-1α in unknown samples is interpolated from a standard curve.

Assay Performance


TABLE 1. Analytical sensitivity of the Erenna® IL-1α Immunoassay Kit1
Lower Limit of Detection 0.25 pg/mL
Lower Limit of Quantification2 0.39 pg/mL
Upper Limit of Quantification 100 pg/mL
Low-end CV% Range 2 - 19%
Low-end CV% Average 7%
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-1α] in a healthy reference


FIGURE 1. [IL-1α] in EDTA plasma from 10 healthy donors (5 samples < LLoQ), with median and interquartile range. The Erenna® IL-1α Immunoassay reliably quantifies IL-1α in healthy subjects, who have a median [IL-1α] of 1.06 pg/mL that is well above the detection limit of 0.25 pg/mL.

FIGURE 2. Erenna® IL-1α Immunoassay Evaluation Reagent 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

IL-1α and IL-1β are pro-inflammatory cytokines involved in immune defense against infection, and are part of the IL-1 superfamily of cytokines. Both IL-1α and IL-1β are produced by macrophages, monocytes and dendritic cells. IL-1 is involved in various immune responses with a primary role in inflammation, making it a target for Rheumatoid Arthritis (RA). IL-1α and IL-1β are produced as precursor peptides, which are proteolytically processed and released in response to cell injury, and thus induce apoptosis. IL-1β production in peripheral tissue has also been associated with hyperalgesia (increased sensitivity to pain) associated with fever.


TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-1 alpha
UniProtKB/Swiss-Prot ID: P01583
Protein-protein interaction database: P01583
Alternative Names: Hematopoietin-1
Gene Names: IL1A
Synonym Gene Names: IL1F1
Function: Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells.
Biological Processes: anti-apoptosis, apoptosis, cell proliferation, cellular response to heat, cytokine-mediated signaling pathway, fever, immune response, negative regulation of cell proliferation, positive regulation of angiogenesis, positive regulation of cell division, positive regulation of cytokine secretion, positive regulation of interleukin-2 biosynthetic process, positive regulation of mitosis, positive regulation vascular endothelial growth factor production, response to copper ion
Molecular Function: copper ion binding, cytokine activity, interleukin-1 receptor binding

* Source Info from www.uniprot.org.


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