Simple and efficient identification of chromatin modifying complexes and characterization of complex composition

Jeong Heon Lee, David Skalnik

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Affinity purification and mass spectrometry analysis have been used to identify and characterize protein complexes. Wdr82-associated chromatin modifying complexes were purified by single-step FLAG affinity purification from human cells induced to express FLAG-tagged Wdr82. Purified proteins were analyzed by SDS-PAGE and specific protein bands were identified by mass spectrometry. Subsequently, purified proteins were fractionated on sucrose gradient equilibrium centrifugation to determine overall composition of each identified complex. We describe here simple and efficient approaches for the identification of chromatin modifying complexes and subsequent characterization of complex composition.

Original languageEnglish (US)
Title of host publicationGene Regulation
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages289-298
Number of pages10
ISBN (Print)9781627032834
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume977
ISSN (Print)1064-3745

Keywords

  • Complex composition
  • FLAG affinity purification
  • Mass spectrometry
  • Protein complex
  • Sucrose gradient equilibrium centrifugation
  • Wdr82

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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