Preparation of disease-related protein assemblies for single particle electron microscopy

A. Cameron Varano, Naoe Harafuji, William Dearnaley, Lisa Guay-Woodford, Deborah F. Kelly

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Electron microscopy (EM) is a rapidly growing area of structural biology that permits us to decode biological assemblies at the nanoscale. To examine biological materials for single particle EM analysis, purified assemblies must be obtained using biochemical separation techniques. Here, we describe effective methodologies for isolating histidine (his)-tagged protein assemblies from the nucleus of disease-relevant cell lines. We further demonstrate how isolated assemblies are visualized using single particle EM techniques and provide representative results for each step in the process.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages185-196
Number of pages12
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1647
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Cameron Varano, A., Harafuji, N., Dearnaley, W., Guay-Woodford, L., & Kelly, D. F. (2017). Preparation of disease-related protein assemblies for single particle electron microscopy. In Methods in Molecular Biology (pp. 185-196). (Methods in Molecular Biology; Vol. 1647). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7201-2_12