Magnetic Bead-Based Isolation of Exosomes
Exosomes are here defined as extracellular vesicles (EVs) in the approximate size range of 30–100 nm in diameter, and are observed in most body fluids containing typical exosomal markers such as CD9, CD63, and CD81. Potential subpopulations of exosomes can be captured by targeting these markers using magnetic beads. Magnetic beads are versatile tools for exosome isolation and downstream analysis. Here, we describe the workflow of immuno magnetic isolation and analysis of exosomes by flow cytometry, Western immunoblotting, and electron microscopy.
Key wordsExtracellular vesicles Exosomes Magnetic bead isolation Dynabeads Electron microscopy
Bente Kierulf, Anette Kullman, and Malin Karlsson (Life Technologies) for expert advice and critically reading the manuscript, Sasha Vlassov and Mu Li (Life Technologies) for NanoSight® data, Antje Hoenen and Norbert Roos (University of Oslo, Norway) for technical assistance in electron microscopy.
- 8.Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol 3:3–22Google Scholar
- 13.Lewis GD, Metcalf TG (1988) Polyethylene glycol precipitation for recovery of pathogenic viruses, including Hepatitis A virus and human Rotavirus, from oyster, water, and sediment samples. Appl Environ Microbiol 4:5Google Scholar
- 14.Yoshioka Y, Konishi Y, Kosaka N, Katsuda T, Kato T, Ochiya T (2013) Comparative marker analysis of extracellular vesicles in different human cancer types. J Extracell Vesicles 2Google Scholar