EYRAplex: Human Th1/Th2 (7‑plex)

Standard curves for EYRAplex: Human Th1/Th2 (7-plex).

EYRAplex: Human Th1/Th2 (7‑plex)

5410-1EY-1

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Components

Capture mAbCapture bead mix: Human Th1/Th2 (7-plex)
Detection mAbDetection mAb mix: Human Th1/Th2 (7-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: Th1/Th2 (7-plex) is intended for the quantification of human IFN-γ, IL-2, IL-4, IL-6, IL-10, IL-12 (p70), and TNF-α in serum, plasma, and cell culture supernatants. The assay is designed for analysis using Mabtech EYRA™ and is also compatible with many flow cytometry platforms. 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 Th1/Th2 (7-plex)
ApplicationMultiplex
AnalyteIFN-γ, IL-2, IL-4, IL-5, IL-6, IL-12 (p70), 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)Intra-assay CV(%)Inter-assay CV(%)
IFN-γ0.13.7<5%<15%
IL-20.11.7<5%<15%
IL-40.29.9<5%<15%
IL-50.22.2<5%<15%
IL-60.22.6<5%<15%
IL-12 (p70)0.56.6<5%<15%
TNF-α1.018.5<5%<15%

Indicative performance from data from Mabtech EYRA™ is shown.

Average Recovery (%) in
AnalyteHeparin plasmaRA plasmaSerum
IFN-γ857988
IL-281102101
IL-49796128
IL-58410381
IL-68999100
IL-12 (p70)-86-
TNF-α9410072

Indicative performance from data from Mabtech EYRA™ is shown.

- Not determined

Standard curves for EYRAplex: Human Th1/Th2 (7-plex).

Representative standard curves for EYRAplex: Human Th1/Th2 (7-plex).

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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-5

Analyte description

Interleukin 5 (IL-5) is a pleiotropic cytokine primarily produced by T and NK cells.  IL-5 specifically controls the survival, differentiation, and chemotaxis of eosinophils. But initially, IL-5 was characterized by its ability to support the growth and terminal differentiation of B cells.

Alternative namesInterleukin-5, IL-5, IL5, BCGF-2
Cell typeT cell, Th2, NK cell

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-12 (p70)

Analyte description

Interleukin 12 (IL-12) is, when biologically active, a heterodimer (p70) consisting of two covalently linked subunits, p35 and p40. The p40 subunit is shared with IL-23. IL-12 is produced by antigen-presenting cells, specifically dendritic cells, and macrophages. IL-12 is a proinflammatory cytokine that promotes Th1-type responses by inducing IFN-γ production and enhancing the proliferation and cytotoxicity of NK and T cells.

Alternative namesInterleukin 12, IL-12, IL12
Cell typeMonocyte/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