Efficient sequential recovery of nucleolar macromolecular components

Baoyan Bai, Marikki Laiho

Research output: Contribution to journalArticle

Abstract

Efficient extraction and accurate quantification of nucleolar macromolecules are critical for in vitro analysis, especially for studying RNA, DNA, and protein dynamics under identical conditions. There is presently no single method that efficiently and simultaneously isolates these three macromolecular constituents from purified nucleoli. We have developed an optimized method, which without evident loss, extracts, and solubilizes protein recovered from a single sample following TRIzol isolation of RNA and DNA. The solubilized protein can be accurately quantified by protein bicinchoninic acid assay and assessed by polyacrylamide gel electrophoresis. We have successfully applied this approach to extract and quantify all three nucleolar components, and to study nucleolar protein responses after actinomycin D treatment.

Original languageEnglish (US)
Pages (from-to)3044-3048
Number of pages5
JournalProteomics
Volume12
Issue number19-20
DOIs
StatePublished - Oct 2012

Keywords

  • Cell biology
  • DNA
  • Macromolecular
  • Nucleolus
  • RNA
  • TRIzol

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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