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From tube to structure: SPA Cryo-EM workflow using apoferritin as an example

Item Type:Article
Title:From tube to structure: SPA Cryo-EM workflow using apoferritin as an example
Creators Name:Diebolder, C.A. and Dillard, R.S. and Renault, L.
Abstract:In this chapter, we present an overview of a standard protocol to achieve structure determination at high resolution by Single Particle Analysis cryogenic Electron Microscopy using apoferritin as a standard sample. The purified apoferritin is applied to a glow-discharged support and then flash frozen in liquid ethane. The prepared grids are loaded into the electron microscope and checked for particle spreading and ice thickness. The microscope alignments are performed and the data collection session is setup for an overnight data collection. The collected movies containing two-dimensional images of the apoferritin sample are then processed to obtain a high-resolution three-dimensional reconstruction.
Keywords:Single Particle Analysis, Standard Protocol, Workflow, Cryo-EM, Cryogenic Electron, Microscopy, Sample Preparation, Animals
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Structural proteomics: high-throughput methods
Publisher:Springer / Humana Press
Page Range:229-256
Number of Pages:344
Date:6 May 2021
Official Publication:https://doi.org/10.1007/978-1-0716-1406-8_12
PubMed:View item in PubMed

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