Leveraging MSD Assay for Cytokine Detection and Analysis

By | November 9, 2021

This article will provide an overview of the MSD assay and how it can be used for cytokine detection and analysis in biologic research. The MSD assay is an effective tool for identifying proteins present in cell extracts or serum or quantifying target proteins present in individual cells or tissues. This  potent analytical tool is particularly suited to detectand quantify immunomodulatory factors such as cytokines, chemokines, hormones, growth factors, and neurotransmitters produced by cells.

How to Use MSD Assay for Cytokine Detection?

Protein A-based immune ligand blotting is a simple and sensitive method for detecting cytokines and growth factors in cell extracts and serum. It can also be used to determine whether a protein target is present in individual cells. Thus, the assay is both specific and quantitative, with little background interference from endogenous proteins or other biomarkers.

The protein A-based immune ligand blotting assay combines the use of protein A chromatography with immunoblotting. After purification and concentration of the desired cell extracts and serum, the samples are incubated with protein A immobilized on a nitrocellulose membrane. The method allows the detection of proteins present in the selection and quantitative determination of their quantity.

The meso scale discovery cytokine (Immunostaining Detection Kits) is a patented technique for simultaneous detection and quantitation of cytokines, chemokines, hormones, growth factors, and neurotransmitters present in cells or samples. In addition, the assay allows for the quantitative determination of proteins present in cell extracts or serum samples; it is used to detect a wide range of these mediators. MSD assay kits have been developed for various cell types and cell culture systems, each with its  specific requirements.

Due to its very low background interference, the MSD assay is more sensitive than other assays or immune analyses used in immunological or biomedical research.. It only requires one incubation (usually overnight) and can be performed at room temperature. As a result, MSD has become an indispensable tool for biomedical researchers.

Leveraging Detection and Analysis Process

As the MSD assay allows detection of several cytokines, chemokines, neurotransmitters, and neuropeptides simultaneously in a single sample (i.e., one example is used per sample type), it has become increasingly popular for multitarget profiling in cell-based assays (i.e., in a single sample one would perform a cDNA microarray to profile all of the targets in a particular arrayed gene set, and determine target-specific mRNA levels, at the same time as completing multiple immunostainings to simultaneously profile various cytokines, chemokines, neuropeptides or growth factors).

In immunohistochemistry, the MSD assay can visualize multiple targets on a slide by applying different antibodies at different points during staining. Unlike immunohistochemistry techniques such as formalin fixation and heat treatment over-expressing antibodies for several days before staining, MSD assay can be used with any antibody(s) with minimal delay. Thus, it is considered an ideal technique for the detection of several targets in immunohistochemistry. Furthermore, it can be performed after sample collection, although optimal results are achieved 48-72 hours after cell isolation.


The MSD assay is extremely useful for detecting multiple targets simultaneously in tissues or cells, whereas many other analytical techniques are limited to seeing only one target at a time. It has provided population profiling, simultaneously quantifying multiple cytokines in biopsied tissue samples from human subjects. These data could then be used for epidemiological studies or diagnostic testing.

Moreover, MSD assay can also  detect and quantify several proteins simultaneously in cell culture supernatants and cell cultures.

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