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Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
Published on: November 9, 2017
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Method to quantify cytokines and chemokines in mouse brain tissue using Bio-Plex multiplex immunoassays
Monica Manglani1, Rejane Rua1, Amy Hendricksen2
1Viral Immunology and Intravital Imaging Section, National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
Methods (San Diego, Calif.)
|February 12, 2019
Summary
This study details a method for analyzing 33 inflammatory mediators in mouse brain tissue using multiplex bead-based immunoassays. This technique enables precise quantification of multiple analytes from limited tissue samples.
Area of Science:
- Neuroscience
- Immunology
- Biochemistry
Background:
- Quantifying inflammatory mediators in small tissue samples is challenging.
- Multiplex assays offer efficient analysis of multiple analytes simultaneously.
- Luminex xMAP technology (Bio-Plex) is suitable for high-throughput analysis.
Purpose of the Study:
- To describe a protocol for preparing mouse brain tissue for multiplex immunoassay.
- To enable simultaneous quantification of 33 inflammatory cytokines and chemokines.
- To validate the use of Bio-Plex Pro Mouse Chemokine Panel 33-Plex for brain tissue analysis.
Main Methods:
- Preparation of mouse brain tissue.
- Utilizing Bio-Plex Pro Mouse Chemokine Panel 33-Plex, a Luminex xMAP-based assay.
- Performing multiplex bead-based immunoassay for analyte quantification.
Main Results:
- Successful simultaneous analysis of 33 inflammatory mediators.
- Demonstration of a reliable method for small-volume tissue analysis.
- Quantification of cytokines and chemokines in mouse brain homogenates.
Conclusions:
- The described protocol effectively prepares mouse brain tissue for multiplex analysis.
- The Bio-Plex Pro Mouse Chemokine Panel 33-Plex is a valuable tool for studying neuroinflammation.
- This method facilitates comprehensive profiling of inflammatory mediators in neurological research.
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