EYRAplex: Human In vitro cytokine release (8‑plex)

Standard curves for EYRAplex: Human In vitro cytokine release (8-plex).

EYRAplex: Human In vitro cytokine release (8‑plex)

5460-1EY-1

$1,800

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Components

Capture mAbCapture bead mix: Human In vitro cytokine release (8-plex)
Detection mAbDetection mAb mix: Human In vitro cytokine release (8-plex)
StandardStandard mix A: EYRAplex, human
Fluorophore conjugateStreptavidin-PE
Buffers/SolutionsAssay diluent: EYRAplex
Streptavidin-PE diluent
Wash buffer concentrate
Also provided96-well plate (black)
Adhesive plate covers
Black plate lid

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Product specifications

Intended use

EyraPlex: In vitro cytokine release (8-plex) is intended for the quantification of human GM-CSF, IFN-γ, IL-2, IL-4, IL-6, IL-8 (CXCL8), IL-10, and TNF-α in serum, plasma, and cell culture supernatants. The assay is designed for analysis using Mabtech EYRA™, but can also be used in flow cytometry. For research use only. Not for use in diagnostic procedures.

Introduction

EYRAplex is a magnetic bead-based multiplex assay for the simultaneous detection and quantification of multiple analytes in a single sample. The magnetic beads have been dyed with different concentrations of two fluorophores, resulting in distinct bead populations with unique bead IDs. Each bead population has been conjugated with a specific monoclonal antibody, allowing specific binding to its corresponding analyte. Multiplex analysis is enabled by combining several bead populations that are supplied as a capture bead mix.

In EYRAplex, the capture bead mix is added to wells in a 96-well plate. Samples and standards are then added. During incubation, the capture antibodies bind to analytes present in the sample. Detection occurs in two steps. First, a biotin-labeled monoclonal antibody mix (detection mAb mix) is added. These detection antibodies bind to the captured analytes. Streptavidin-PE is then added, binding to the biotin and generating a fluorescent signal proportional to the analyte concentration.

The fluorescent signal from each bead ID determines the analyte, while the streptavidin-PE signal reflects its quantity. Analytes are quantified by comparing the signals to standard curves.

Serum/Plasma samples

EYRAplex is designed to minimize interference from heterophilic antibodies and rheumatoid factor (RF), which can lead to false-positive results in immunoassays. The assay has been validated using serum and plasma samples from healthy donors, as well as samples containing elevated RF levels.

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Product details

ProductEYRAplex: Human In vitro cytokine release (8-plex)
ApplicationMultiplex, Flow cytometry
AnalyteGM-CSF, IFN-γ, IL-2, IL-4, IL-6, IL-8 (CXCL8), IL-10, TNF-α
ReactivityHuman

Shipping and Storage

Shipping

Shipped with cold packs.

Storage

Store at 4-8 °C upon receipt.

AnalyteLOD (pg/ml)LLOQ (pg/ml)ULOQ (pg/ml)Intra-assay CV(%)Inter-assay CV(%)
GM-CSF0.10.754047
IFN-γ0.10.861047
IL-20.26.1147046
IL-40.23.3240057
IL-60.13.3243045
IL-8 (CXCL8)0.536.0434057
IL-100.11.284036
TNF-α0.217.5141336

Intra-assay and inter-assay precision were evaluated using the assay standard at three concentrations (high, medium, low) in Assay diluent: EYRAplex. CV values are based on several independent experiments, using eight replicates per condition.

Average recovery (%) in
AnalyteHeparin plasmaCitrate plasmaEDTA plasmaSerum
GM-CSF79897790
IFN-γ74776684
IL-291929474
IL-4829893*
IL-683928688
IL-8 (CXCL8)9010594111
IL-1076768375
TNF-α64818162

Spike recovery was assessed in different matrices from healthy donors using the assay standard at three concentrations (high, medium, low) and unspiked controls, with four replicates per condition. Several independent experiments were performed, and the average recovery was calculated relative to spiked Assay diluent: EYRAplex.

*IL-4: Not recommended

Average recovery (%) in
AnalyteRA plasma
GM-CSF79
IFN-γ79
IL-2102
IL-496
IL-699
IL-8 (CXCL8)97
IL-1091
TNF-α100

Spike recovery in RA plasma (high rheumatoid factor) was evaluated during assay development. Indicative results shown.

Standard curves for EYRAplex: Human In vitro cytokine release (8-plex).

Representative standard curves for EYRAplex: Human In vitro cytokine release (8-plex).

Tutorial, April 11, 2025

Step-by-step guide to EYRAplex

New to magnetic bead-based multiplex assays? No worries! We've got you covered with this in-depth tutorial explaining each step in the assay as well as tips and tricks along the way.
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GM-CSF

Analyte description

Granulocyte macrophage-colony stimulating factor (GM-CSF) can be secreted by T cells, macrophages, endothelial cells, and fibroblasts. GM-CSF stimulates the survival and functional activities of myeloid, that is, monocytes, macrophages, DCs, neutrophils, and eosinophils. It also stimulates the differentiation and proliferation of hematological progenitors.

Alternative namesGranulocyte macrophage-colony stimulating factor, GM-CSF, CSF2
Cell typeT cell

IFN-γ

Analyte description

Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and cytotoxic T cells is a hallmark of the Th1-type phenotype. Thus, high-level production of IFN-γ is typically associated with effective host defense against intracellular pathogens.

Alternative namesInterferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg
Cell typeT cell, Tc, Th1, NK cell

IL-2

Analyte description

Interleukin 2 (IL-2) is predominantly produced by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK cells. After antigen stimulation, the cytokine also promotes the differentiation of T cells into effector and memory T cells. Therefore IL-2 plays an important role in the body's response to infection. IL-2 is also key to tolerance, as it promotes the differentiation of certain immature T cells into regulatory T cells in the thymus, ultimately preventing autoimmune diseases. 

Alternative namesInterleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor
Cell typeT cell, Th1, Th2, Th17, Tfh

IL-4

Analyte description

Interleukin 4 (IL-4) is produced primarily by Th2 cells, mast cells, eosinophils, and basophils. IL-4 was first identified to costimulate B cell growth; the cytokine is essential for B cell secretion of IgE and augments the production of IgG1 in vivo. In addition to regulating B cell growth and immunoglobulin secretion, IL-4 also affects T cells. In vitro, IL-4 promotes T cell growth and can induce cytolytic T cell activity. IL-4 also induces the differentiation of naive CD4 T cells into Th2 cells, which are characterized by their capacity to secrete the cytokines IL-4, IL-5, and IL-10 upon activation, while simultaneously inhibiting the generation of Th1 cells.

Alternative namesInterleukin 4, IL-4, IL4, BCGF-1, BSF-1
Cell typeT cell, Th2, Treg

IL-6

Analyte description

Interleukin 6 (IL-6) is a pleiotropic cytokine produced by many different cell types and plays a role in a wide range of functions, such as immune responses, acute-phase reactions, and hematopoiesis. Among other things, it augments antibody production from activated B cells in vitro.

Alternative namesInterleukin 6, IL-6, IL6, IFB-B502, BSF-2, BCDF
Cell typeB cell, Monocyte/MΦ, mDC

IL-8 (CXCL8)

Analyte description

Interleukin 8 (IL-8) is a proinflammatory cytokine produced by different types of cells, including monocytes, macrophages, and endothelial cells. Inflammatory stimuli, such as LPS, strongly induce IL-8 production by monocytes.

Alternative namesInterleukin-8, IL-8, IL8, CXCL8
Cell typeT cell, Treg, Tfh, Monocyte/MΦ, mDC

IL-10

Analyte description

Interleukin 10 (IL-10) is a pleiotropic cytokine produced by activated T cells and monocytes. It can modulate the functions of immune responses in both a stimulatory and an inhibitory way. IL-10 is thereby involved in the regulation of immune reactions and inflammatory responses.

Alternative namesInterleukin-10, IL-10, IL10
Cell typeT cell, Treg, Monocyte/MΦ, mDC

TNF-α

Analyte description

Tumor necrosis factor (TNF), also known as TNF-α, is produced by many different cell types, e.g., monocytes, macrophages, T cells, and B cells. Among the many effects of TNF-α are protection against bacterial infection, cell growth modulation, immune system regulation, and involvement in septic shock.

Alternative namesTumor necrosis factor-α, TNF-α, TNF-alpha, TNF-a, TNFa, Tumor necrosis factor-alpha, TNF
Cell typeT cell, Tc, Th1, Th2, Th17, Tfh, Monocyte/MΦ