Biomarker Analytes: GM-CSF, IFNγ, IL-1 β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p70), IL-13, IL-17A, IL-23 and TNFα.
Biomarker Research Areas: Immune response, cardiovascular disease, diabetes, neurological disorders and cancer.
Multiplex Assay Overview: The assay service provided by Eve Technologies on their Human High Sensitivity T-Cell Discovery Array 14-Plex facilitates research on immune response, cardiovascular disease, diabetes, neurological disorders and cancer. This Discovery Assay® measures fourteen biomarkers simultaneously in a single microwell. The assay is designed for extracellular protein analysis in nearly all types of biological samples.
Background Information: Cytokines are immunomodulatory polypeptides that play key roles in both adaptive and innate immune responses. A generic term, “cytokines” includes myokines (produced by muscle cells), lymphokines (produced by activated Th cells), interleukins (acting as mediators between T cells) and chemokines (responsible for T-cell migration). One of the regulatory mechanisms of the immune system, cytokines act at the recognition, activation, or effector phases of an immune response, modulating the development and functional activities of the subtypes of T cells, B cells and myeloid cells. Consequently, research involving cytokines plays a significant role in achieving a deeper understanding of the immune system and its multi-faceted response to most antigens, especially those responses that make up the inflammatory process.
Low levels of inflammation are involved in many clinical and sub-clinical disease states, such as autoimmune disease, cardiovascular disease, diabetes, neurological disorders and cancer. Measuring picogram levels of cytokines, therefore, is critical for understanding the pathogenesis of these diseases.
Intended Sample Type: Human serum or plasma (EDTA recommended), cell cultures, tissue homogenates and others. Due to a wide range of sample conditions, a pilot run is recommended for cell cultures, tissue homogenates, and other sample types not listed.
Special Sample Preparation: ALL samples must be centrifuged prior to aliquoting. The aliquot should be drawn from the middle of the sample volume. Samples that have a high amount of contaminants, cellular debris, lipids, or coagulation can result in a volume loss as volume is being displaced by this debris and they can clog the filters. Highly turbid or coagulated samples that clog the filters will be partially filtered by piercing the clogged membrane.
All samples will be diluted 2-fold by Eve Technologies, as recommended by Millipore for this assay.
Usage Statement: Unless otherwise stated our products and services are intended for research use only.
Detection Method: Multiplex Immunoassay analyzed with a BioPlex 200
Human High Sensitivity T-Cell Discovery Array 14-Plex Assay Kit Source: Millipore MILLIPLEX
|Sensitivity, Variation, & Accuracy
*Note: %CV is an appropriate measure of quality only for sample that have significant detectable concentrations for a particular analyte. Low concentration / low signal samples can often have a much higher %CV as the signals are at the limitations of the assay and fall within a flat portion of the curve.
% recovery is comparing the observed concentration to the expected concentration of samples spiked with the standard curve cocktail at various concentrations.
|GM-CSF||0.35||< 5||< 15||101|
|IFNγ||0.48||< 5||< 20||101|
|IL-1β||0.14||< 5||< 15||98|
|IL-2||0.19||< 5||< 15||103|
|IL-4||1.12||< 5||< 15||103|
|IL-5||0.12||< 5||< 20||101|
|IL-6||0.11||< 5||< 20||107|
|IL-8||0.13||< 5||< 15||103|
|IL-10||0.56||< 5||< 20||104|
|IL-12 (p70)||0.15||< 6||< 15||100|
|IL-13||0.23||< 5||< 20||103|
|IL-17A||0.33||< 5||< 20||106|
|IL-23||3.25||< 5||< 20||100|
|TNFα||0.16||< 5||< 15||103|
|Normal Reference Range||TBD|
Multiplex Assay Methods
Below is a methods description that is often used in publications.
Using a methods description that credits Eve Technologies in a publication will result in a 10% discount on your next order!
Multiplex analysis of cytokines
In this study we quantified 14 cytokine/ chemokine/ growth factor biomarkers simultaneously by using a Discovery Assay® called the Human High Sensitivity T-Cell Discovery Array 14-Plex (Eve Technologies Corp, Calgary, AB, Canada). The multiplex assay was performed at Eve Technologies by using the Bio-Plex™ 200 system (Bio-Rad Laboratories, Inc., Hercules, CA, USA), and a Milliplex Human High Sensitivity T-Cell panel (Millipore, St. Charles, MO, USA) according to their protocol. The 14-plex consisted of GM-CSF, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p70), IL-13, IL-17A, IL-23, and TNFα. The assay sensitivities of these markers range from 0.11 – 3.25 pg/mL. Individual analyte values and other assay details are available on Eve Technologies’ website or in the Milliplex protocol.