GraFix: sample preparation for single-particle electron cryomicroscopy

被引：399

作者：

Kastner, Berthold ^{[1
]}

Fischer, Niels ^{[2
]}

Golas, Monika Mariola ^{[2
]}

Sander, Bjoern ^{[2
]}

Dube, Prakash ^{[2
]}

Boehringer, Daniel ^{[2
]}

Hartmuth, Klaus ^{[1
]}

Deckert, Jochen ^{[1
]}

Hauer, Florian ^{[2
]}

Wolf, Elmar ^{[1
]}

Uchtenhagen, Hannes ^{[2
]}

Urlaub, Henning ^{[3
]}

Herzog, Franz ^{[5
]}

Peters, Jan Michael ^{[5
]}

Poerschke, Dietmar ^{[4
]}

Hrmann, Reinhard Lu ^{[1
]}

Stark, Holger ^{[2
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept Cellular Biochem, D-37077 Gottingen, Germany

[2] Max Planck Inst Biophys Chem, Electron Cryomicroscopy Grp 3D, D-37077 Gottingen, Germany

[3] Max Planck Inst Biophys Chem, Mass Spectrometry Grp, D-37077 Gottingen, Germany

[4] Max Planck Inst Biophys Chem, Biomol Dynam Grp, D-37077 Gottingen, Germany

[5] Res Inst Mol Pathol, A-1030 Vienna, Austria

来源：

NATURE METHODS | 2008年 / 5卷 / 01期

关键词：

D O I：

10.1038/NMETH1139

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：53 / 55

页数：3

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