![]() ![]() 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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