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- Title
GraFix: sample preparation for single-particle electron cryomicroscopy.
- Authors
Kastner, Berthold; Fischer, Niels; Golas, Monika Mariola; Sander, Bjoern; Dube, Prakash; Boehringer, Daniel; Hartmuth, Klaus; Deckert, Jochen; Hauer, Florian; Wolf, Elmar; Uchtenhagen, Hannes; Urlaub, Henning; Herzog, Franz; Peters, Jan Michael; Poerschke, Dietmar; Lührmann, Reinhard; Stark, Holger
- Abstract
We developed a method, named GraFix, that considerably improves sample quality for structure determination by single-particle electron cryomicroscopy (cryo-EM). GraFix uses a glycerol gradient centrifugation step in which the complexes are centrifuged into an increasing concentration of a chemical fixation reagent to prevent aggregation and to stabilize individual macromolecules. The method can be used to prepare samples for negative-stain, cryo-negative-stain and, particularly, unstained cryo-EM.
- Subjects
CRYOMICROSCOPY; MACROMOLECULES; GLYCERIN; ELECTRON microscopy; CELL aggregation
- Publication
Nature Methods, 2008, Vol 5, Issue 1, p53
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth1139