Proteomic analysis of urinary exosomes in cardiovascular and associated kidney diseases by two-dimensional electrophoresis and LC-MS/MS

Methods Mol Biol. 2013:1000:209-20. doi: 10.1007/978-1-62703-405-0_16.

Abstract

Urinary exosomes are membranous vesicles 40-100 nm in size containing proteins that are characteristic of every renal tubule epithelial cell type. In this chapter, we describe a methodology to isolate and analyze urinary exosomes proteome by 2-DE and LC-MS/MS, in the search for biomarkers of vascular and associated kidney diseases. We describe an isolation methodology by serial (ultra)centrifugation steps compatible with 2-DE and LC-MS/MS analysis. Exosome purity is confirmed by electron microscopy and Western blot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / urine
  • Blotting, Western
  • Cardiovascular Diseases / complications*
  • Centrifugation
  • Chromatography, Liquid / methods*
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Exosomes / metabolism*
  • Exosomes / ultrastructure
  • Humans
  • Kidney Diseases / complications
  • Kidney Diseases / pathology
  • Kidney Diseases / urine*
  • Microscopy, Electron
  • Proteomics / methods*
  • Specimen Handling
  • Tandem Mass Spectrometry / methods*

Substances

  • Biomarkers