![]() Twomey EC, Yelshanskaya MV, Grassucci RA, Frank J, Sobolevsky AI (2016) Elucidation of AMPA receptor-stargazin complexes by cryo-electron microscopy. Kulig W, Tynkkynen J, Javanainen M, Manna M, Rog T, Vattulainen I, Jungwirth P (2014) How well does cholesteryl hemisuccinate mimic cholesterol in saturated phospholipid bilayers? J Mol Model 20(2):2121. Hauer F, Gerle C, Fischer N, Oshima A, Shinzawa-Itoh K, Shimada S, Yokoyama K, Fujiyoshi Y, Stark H (2015) GraDeR: membrane protein complex preparation for single-particle cryo-EM. Gewering T, Januliene D, Ries AB, Moeller A (2018) Know your detergents: a case study on detergent background in negative stain electron microscopy. Springer New York, New York, NY, pp 21–82. In: Wolkers WF, Oldenhof H (eds) Cryopreservation and freeze-drying protocols. J Electron Microsc Tech 7(1):29–33įahy GM, Wowk B (2015) Principles of cryopreservation by vitrification. Īebi U, Pollard TD (1987) A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. Acta Crystallogr D Struct Biol 74(Pt 6):560–571. ĭrulyte I, Johnson RM, Hesketh EL, Hurdiss DL, Scarff CA, Porav SA, Ranson NA, Muench SP, Thompson RF (2018) Approaches to altering particle distributions in cryo-electron microscopy sample preparation. Li X, Mooney P, Zheng S, Booth CR, Braunfeld MB, Gubbens S, Agard DA, Cheng Y (2013) Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM. This protocol outlines a quick and straightforward procedure for screening and determining the structure of a membrane protein of interest under biologically relevant conditions. Here we describe preparation of negative staining and cryo-EM grids and downstream data collection of membrane proteins in detergent, by far the most common solubilization agent. Specimen preparation remains the bottleneck of most cryo-EM research projects, with membrane proteins representing particularly challenging targets of investigation due to their universal requirement for detergents or other solubilizing agents. ![]() Especially in the context of membrane proteins, this technique has allowed researchers to obtain structural information at a previously unattainable level of detail. Cryo-electron microscopy (cryo-EM) is a powerful tool for investigating the structure of macromolecules under near-native conditions.
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