Magnetic bead-based isolation of exosomes

Methods Mol Biol. 2015:1218:465-81. doi: 10.1007/978-1-4939-1538-5_27.

Abstract

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.

MeSH terms

  • Biomarkers / metabolism
  • Blotting, Western
  • Cell Fractionation / methods*
  • Cell Line, Tumor
  • Exosomes / chemistry*
  • Exosomes / metabolism
  • Exosomes / ultrastructure
  • Flow Cytometry
  • Gene Expression
  • Humans
  • Immunomagnetic Separation / methods*
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / pathology
  • Jurkat Cells
  • Microscopy, Electron
  • Microspheres
  • Tetraspanin 28 / metabolism
  • Tetraspanin 29 / metabolism
  • Tetraspanin 30 / metabolism

Substances

  • Biomarkers
  • CD63 protein, human
  • CD81 protein, human
  • CD9 protein, human
  • Tetraspanin 28
  • Tetraspanin 29
  • Tetraspanin 30