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.
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Acknowledgments
This work was supported by grants from FIS CP09/229, FIS PI11/01401, and FIS PI08/0970.
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Zubiri, I., Vivanco, F., Alvarez-Llamas, G. (2013). Proteomic Analysis of Urinary Exosomes in Cardiovascular and Associated Kidney Diseases by Two-Dimensional Electrophoresis and LC–MS/MS. In: Vivanco, F. (eds) Vascular Proteomics. Methods in Molecular Biology, vol 1000. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-405-0_16
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DOI: https://doi.org/10.1007/978-1-62703-405-0_16
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Publisher Name: Humana Press, Totowa, NJ
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