Actions for Long shelf-life streptavidin support-films suitable for electron microscopy of biological macromolecules [electronic resource].

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Long shelf-life streptavidin support-films suitable for electron microscopy of biological macromolecules [electronic resource].

Published
Washington, D.C. : United States. Dept. of Energy, 2016.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
Physical Description
pages 238-244 : digital, PDF file
Additional Creators
Lawrence Berkeley National Laboratory, United States. Department of Energy, National Institutes of Health (U.S.), and United States. Department of Energy. Office of Scientific and Technical Information
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Summary
We describe a rapid and convenient method of growing streptavidin (SA) monolayer crystals directly on holey-carbon EM grids. As expected, these SA monolayer crystals retain their biotin-binding function and crystalline order through a cycle of embedding in trehalose and, later, its removal. This fact allows one to prepare, and store for later use, EM grids on which SA monolayer crystals serve as an affinity substrate for preparing specimens of biological macromolecules. In addition, we report that coating the lipid-tail side of trehalose-embedded monolayer crystals with evaporated carbon appears to improve the consistency with which well-ordered, single crystals are observed to span over entire, 2 μm holes of the support films. Randomly biotinylated 70S ribosomes are used as a test specimen to show that these support films can be used to obtain a high-resolution cryo-EM structure
Report Numbers
E 1.99:1378360
Subject(s)
Note
Published through SciTech Connect.
06/15/2016.
"ark:/13030/qt9000k1q5"
": S1047847716301150"
Journal of Structural Biology 195 2 ISSN 1047-8477 AM
Bong-Gyoon Han; Zoe Watson; Hannah Kang; Arto Pulk; Kenneth H. Downing; Jamie Cate; Robert M. Glaeser.
Funding Information
AC02-05CH11231
R01-GM083039
P01-GM051487
R01-GM065050
View MARC record | catkey: 23497880