Tissue levels of selected cytokines and chemokines were measured by multiplex ELISA according to the manufacturer's instructions (V-Plex MSD Multi-spot system, Meso Scale Discovery, Rockville, MD). Samples were loaded in 96-well plates pre-coated with target analyte antibodies for two hours at room temperature. Plates were washed, detection antibodies containing electrochemiluminescent tags added, plates were again washed, and a voltage applied to the plates resulted in emission of light by the captured labels. The intensity of light emitted allowed a quantitative measure of analyte concentrations in the samples using a standard curve. Initial assays of a subset of samples measured IL-6, KC/Gro, IL-1β, IFN-γ, IL-10, IL-12, and TNF-α, with only IL-6, KC/Gro, IL-1β, and IFN-γ measured in subsequent assays. Levels of IL-10, IL-12, and TNF-α were below the detection limit of the assay in all of the initial samples tested.
V plex msd multi spot system
Manufactured by Mesoscale
The V-Plex MSD Multi-spot system is a laboratory equipment used for high-throughput multiplex assays. It is designed to simultaneously detect and quantify multiple analytes in a single sample. The system utilizes Mesoscale's proprietary electrochemiluminescence technology to enable sensitive and accurate measurement of proteins, peptides, and other biomolecules.
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2 protocols using v plex msd multi spot system
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Multiplex ELISA for Cytokine Profiling
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Multiplex ELISA for Cytokine Profiling
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Tissue levels of selected cytokines and chemokines were measured by multiplex ELISA according to the manufacturer's instructions (V-Plex MSD Multi-spot system, Meso Scale Discovery, Rockville, MD). Samples were loaded in 96-well plates pre-coated with target analyte antibodies for two hours at room temperature. Plates were washed, detection antibodies containing electrochemiluminescent tags added, plates were again washed, and a voltage applied to the plates resulted in emission of light by the captured labels. The intensity of light emitted allowed a quantitative measure of analyte concentrations in the samples using a standard curve. Initial assays of a subset of samples measured IL-6, KC/Gro, IL-1β, IFN-γ, IL-10, IL-12, and TNF-α, with only IL-6, KC/Gro, IL-1β, and IFN-γ measured in subsequent assays. Levels of IL-10, IL-12, and TNF-α were below the detection limit of the assay in all of the initial samples tested.
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