Abstract
Exosomes are an intriguing class of nanosized vesicles (~30–150 nm in diameter) released by all cell types for intercellular communication and also for cellular metabolic waste removal to maintain cellular homeostasis. Exosomes secreted by cancer cells play an important role in supporting tumor growth and metastasis by communicating with other cells in the tumor microenvironment and distant sites. Several studies have reported that the exosomes secreted by cancer cells show distinct characteristics, including size, cargo, and surface proteins from the normal cells, and can be used as important biomarkers for diagnosis and prognosis for various cancer types. Exosomes represent many distinct biochemical and morphological characteristics than other -extracellular vesicles (EVs), including their size and surface proteins. Understanding the functional role of exosomes requires specific methods for their characterization to distinguish them from other EV and non-EV structures. Transmission electron microscopy with the immunogold labeling method allows direct detection of exosomes based on their size and specific surface protein. In this chapter, we outlined the required materials and detailed method for immunogold labeling for exosomal surface proteins and size characterization.
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Acknowledgments
The authors acknowledge the Department of Defense (DOD) award W81XWH-19-1-0427 (to GD) and R01DA049267 (to GD). The authors also acknowledge Wake Forest Baptist Comprehensive Cancer Center (WFBCCC) Cellular Imaging Shared Resource supported by NCI (P30CA012197, PI: Dr. Boris Pasche).
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Su, Y., Kumar, A., Deep, G. (2022). Characterization of Exosomal Surface Proteins by Immunogold Labeling. In: Deep, G. (eds) Cancer Biomarkers. Methods in Molecular Biology, vol 2413. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1896-7_18
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DOI: https://doi.org/10.1007/978-1-0716-1896-7_18
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