Abstract
Circulating microRNAs (miRNAs) have been recently detected in extracellular body fluids and proved themselves as promising biomarkers for a broad spectrum of diseases. The techniques to isolate, detect, and characterize extracellular miRNAs vary significantly from report to report. In this chapter we describe a consistent, efficient and highly reproducible protocol for isolation and detection of microvesicles-free circulating miRNA from human blood plasma or cell culture media. Furthermore, since exosomes-free circulating miRNAs are associated with Argonaut proteins (the same proteins to which miRNAs are bound inside the cell), we provide a protocol for immunoprecipitation of AGO2 associated miRNAs.
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Acknowledgments
This work was supported by Dietmar-Hopp Foundation, the Helmholtz Society, and the German Cancer Research Center. We thank Mrs. Anne Langheinz for helping to establish miRNA isolation protocol.
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© 2013 Springer Science+Business Media New York
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Turchinovich, A., Weiz, L., Burwinkel, B. (2013). Isolation of Circulating MicroRNA Associated with RNA-Binding Protein. In: Kosaka, N. (eds) Circulating MicroRNAs. Methods in Molecular Biology, vol 1024. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-453-1_8
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DOI: https://doi.org/10.1007/978-1-62703-453-1_8
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-452-4
Online ISBN: 978-1-62703-453-1
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