title: Proteomic analysis of urinary exosomes in cardiovascular and associated kidney diseases by two-dimensional electrophoresis and LC-MS/MS.
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.
authors.affiliation: Department of Immunology, Instituto de Investigacion Sanitaria Fundacion Jimenez Diaz (IIS-FJD), Madrid, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Zubiri
authors.forename: Fernando
authors.initials: F
authors.lastname: Vivanco
authors.forename: Gloria
authors.initials: G
authors.lastname: Alvarez-Llamas
mesh_terms: D015415:Biomarkers / Q000652:urine; D015153:Blotting, Western; D002318:Cardiovascular Diseases / Q000150:complications*; D002498:Centrifugation; D002853:Chromatography, Liquid / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D055354:Exosomes / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007674:Kidney Diseases / Q000150:complications / Q000473:pathology / Q000652:urine*; D008854:Microscopy, Electron; D040901:Proteomics / Q000379:methods*; D013048:Specimen Handling; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers