GraFix: sample preparation for single-particle electron cryomicroscopy.

Kastner, Berthold; Fischer, Niels; Golas, Monika Mariola; Sander, Bjoern; Dube, Prakash; Boehringer, Daniel; Hartmuth, Klaus; Deckert, Jochen et al. · Nat Methods · 2008

basic_science · Level V

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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.

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