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Methods for Extraction of RNA, Proteins, or Protein Complexes from Subcellular Compartments of Eukaryotic Cells

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mRNA Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1648))

Abstract

Eukaryotic organelles or subcellular compartments can be selectively isolated based on their physical density and their stability in the presence of nonionic detergents. This chapter describes a protocol for the preparation of cytoplasm, nucleoplasm, and chromatin, in addition to isolation of RNA and proteins from these fractions. Proteins and protein complexes stably associated with chromatin and other high molecular weight nuclear components can be extracted under non-denaturing conditions by enzymatic digestion of RNA and DNA. The chapter also includes a detailed extraction protocol from highly purified nuclei.

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Acknowledgment

We thank Julia Nikolic for critical reading of the manuscript and Douglas L. Black for his support. K-H.Y. was supported by a fellowship from UCLA Broad Stem Cell Research Center’s Training Program.

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Authors and Affiliations

  1. Department of Microbiology, Immunology, and Molecular Genetics, The David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA

    Kyu-Hyeon Yeom & Andrey Damianov

Authors
  1. Kyu-Hyeon Yeom
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  2. Andrey Damianov
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Corresponding authors

Correspondence to Kyu-Hyeon Yeom or Andrey Damianov .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, California, USA

    Yongsheng Shi

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Yeom, KH., Damianov, A. (2017). Methods for Extraction of RNA, Proteins, or Protein Complexes from Subcellular Compartments of Eukaryotic Cells. In: Shi, Y. (eds) mRNA Processing. Methods in Molecular Biology, vol 1648. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7204-3_12

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  • DOI: https://doi.org/10.1007/978-1-4939-7204-3_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7203-6

  • Online ISBN: 978-1-4939-7204-3

  • eBook Packages: Springer Protocols

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