https://3demmethods.i2pc.es/index.php?action=history&feed=atom&title=2025Chen_Detergent 2026-05-24T20:20:57Z Revision history for this page on the wiki MediaWiki 1.44.2 https://3demmethods.i2pc.es/index.php?title=2025Chen_Detergent&diff=5078&oldid=prev 2025-10-16T12:45:47Z

<p>Created page with &quot;== Citation == Chen, B., Harrison, P., Kargas, V., Pollock, N., Ford, R.C., Prince, S.M. and Collins, R.F. 2025. Coming Clean and Avoiding Bubble Trouble–Using Detergents Wisely in the Purification of Membrane Proteins for Cryo-EM Studies. Biomolecules. 15, 9 (2025), 1315. == Abstract == Detergent solubilisation remains the most commonly used but potentially problematic method to extract membrane proteins from lipid bilayers for Cryo-EM studies. Although recent adva...&quot;</p> <p><b>New page</b></p><div>== Citation ==<br /> <br /> Chen, B., Harrison, P., Kargas, V., Pollock, N., Ford, R.C., Prince, S.M. and Collins, R.F. 2025. Coming Clean and Avoiding Bubble Trouble–Using Detergents Wisely in the Purification of Membrane Proteins for Cryo-EM Studies. Biomolecules. 15, 9 (2025), 1315.<br /> <br /> == Abstract ==<br /> <br /> Detergent solubilisation remains the most commonly used but potentially problematic<br /> method to extract membrane proteins from lipid bilayers for Cryo-EM studies. Although<br /> recent advances have introduced excellent alternatives—such as amphipols, nanodiscs<br /> and SMALPs—the use of detergents is often necessary for intermediate steps. In this<br /> paper, we share our experiences working with detergent-solubilised samples within the<br /> modern Cryo-EM structural pipeline from the perspective of an EM specialist. Our aim is<br /> to inform novice users about potential challenges they may encounter. Drawing on specific<br /> examples from a variety of biological membrane systems, including Magnesium channels,<br /> lipopolysaccharide biosynthesis, and the human major facilitator superfamily transporters,<br /> we describe how the intrinsic properties of detergent-extracted samples can affect protein<br /> purification, Cryo-EM grid preparation (including the formation of vitreous ice) and the<br /> reconstitution of proteins into micelles. We also discuss how these unique characteristics<br /> can impact different stages of structural analysis and lead to complications in single-particle<br /> averaging software analysis. For each case, we present our insights into the underlying<br /> causes and suggest possible mitigations or alternative approaches.<br /> <br /> == Keywords ==<br /> <br /> == Links ==<br /> <br /> https://www.mdpi.com/2218-273X/15/9/1315<br /> <br /> == Related software ==<br /> <br /> == Related methods ==<br /> <br /> == Comments ==</div>

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