Membrane Protein Cryo-EM: Cryo-Grid Optimization and Data Collection with Protein in Detergent
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Membrane Protein Cryo-EM : Cryo-Grid Optimization and Data Collection with Protein in Detergent. / Bloch, Magnus; Santiveri, Mònica; Taylor, Nicholas M.I.
Expression, Purification, and Structural Biology of Membrane Proteins. red. / Camilo Perez; Timm Maier. Humana Press, 2020. s. 227-244 (Methods in Molecular Biology, Bind 2127).Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
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TY - CHAP
T1 - Membrane Protein Cryo-EM
T2 - Cryo-Grid Optimization and Data Collection with Protein in Detergent
AU - Bloch, Magnus
AU - Santiveri, Mònica
AU - Taylor, Nicholas M.I.
PY - 2020
Y1 - 2020
N2 - Cryo-electron microscopy (cryo-EM) is a powerful tool for investigating the structure of macromolecules under near-native conditions. Especially in the context of membrane proteins, this technique has allowed researchers to obtain structural information at a previously unattainable level of detail. 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. 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. This protocol outlines a quick and straightforward procedure for screening and determining the structure of a membrane protein of interest under biologically relevant conditions.
AB - Cryo-electron microscopy (cryo-EM) is a powerful tool for investigating the structure of macromolecules under near-native conditions. Especially in the context of membrane proteins, this technique has allowed researchers to obtain structural information at a previously unattainable level of detail. 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. 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. This protocol outlines a quick and straightforward procedure for screening and determining the structure of a membrane protein of interest under biologically relevant conditions.
KW - Cryo-electron microscopy
KW - Data collection
KW - Detergent
KW - Grid preparation
KW - Membrane protein
KW - Negative staining
KW - Structural biology
KW - Vitrification
U2 - 10.1007/978-1-0716-0373-4_16
DO - 10.1007/978-1-0716-0373-4_16
M3 - Book chapter
C2 - 32112326
AN - SCOPUS:85080912616
SN - 978-1-0716-0372-7
T3 - Methods in Molecular Biology
SP - 227
EP - 244
BT - Expression, Purification, and Structural Biology of Membrane Proteins
A2 - Perez, Camilo
A2 - Maier, Timm
PB - Humana Press
ER -
ID: 238002304