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Cryo-EM Vitrification Starter Kit
单粒子电子冷冻显微镜 (cryo-EM) 是一种强大的技术，可以将蛋白质复合物的结构确定到原子分辨率。 溶液中的样品在液态乙烷中快速冷冻，形成一层薄薄的玻璃冰，其中样品颗粒理想地以随机方向均匀分布，距离空气-水界面足够远（如图）。
然而，冷冻电镜的蛋白质样品玻璃化是一个主要瓶颈。 只有结合选择正确的网格类型、玻璃化条件、蛋白质浓度和添加剂，才能在冷冻电镜中获取高质量的数据。
Cryo-EM V-Kit 是一个玻璃化入门套件，提供已成功应用于低温-EM 样品制备的一系列表面活性剂和一系列 Quantifoil Holey 碳膜，以促进寻找两种可溶性物质的最佳玻璃化条件 和膜蛋白。
Full Description
SKU: M-CEM-VK301Categories: Cryo-EM, Cryo-EM Vitrification Detergents, & Surfactants, New Products, Starter Kits
Product Information
- Product Description
- References
- Specifications
The Cryo-EM V-Kit contains 10 different surfactants plus glycerol as cryoprotectant and 4 different types of Quantifoil grids. It is designed as a Vitrification Starter Kit to facilitate the search for the optimum vitrification condition and cryo-EM grid for both soluble and membrane proteins.
[1] Noble et al. (2018) Routine Single Particle CryoEM Sample and Grid Characterization by Tomography. DOI: 10.7554/eLife.34257.
[2] Thonghin et al. (2018) Cryo-electron microscopy of membrane proteins. Methods 147:176.
[3] Drulyte et al. (2018) Approaches to altering particle distributions in cryo-electron microscopy sample preparation. Acta Cryst. D 74:560.
[4] Glaeser et al. (2017) Opinion: hazards faced by macromolecules when confined to thin aqueous films. Biophys Rep 3:1.
[5] Gatsogiannis et al. (2016). Membrane insertion of a Tc toxin in near-atomic detail. Nat. Struct. Mol. Biol. 23:884.
[6] Efremov et al. (2015) Architecture and conformational switch mechanism of the ryanodine receptor. Nature 517:39.
| Surfactants and Cryoprotectants |
Amount |
Conc. |
CMC |
Class |
| Fluorinated Octyl Maltoside (FOM) |
100 μl |
0.41% (w/v) |
0.07% (w/v) |
non-ionic detergent |
| Hexadecyl-trimethyl-ammonium Bromide (CTAB) |
100 μl |
0.34% (w/v) |
0.03% (w/v) |
cationic detergent |
| n-Decyl-ß-D-Maltoside (DM) |
100 μl |
0.87% (w/v) |
0.09% (w/v) |
non-ionic detergent |
| n-Decyl-α-D-Maltoside (DαM) |
100 μl |
0.46% (w/v) |
0.08% (w/v) |
non-ionic detergent |
| n-Dodecyl-ß-D-Maltoside (DDM) |
100 μl |
0.09% (w/v) |
0.01% (w/v) |
non-ionic detergent |
| Sodium Deoxycholate |
100 μl |
1.66% (w/v) |
0.17% (w/v) |
anionic detergent |
| Triton X-100 |
100 μl |
0.15% (w/v) |
0.01% (w/v) |
non-ionic detergent |
| Tween 20 |
100 μl |
1% (w/v) |
0.01% (w/v) |
non-ionic detergent |
| CHAPSO |
100 μl |
2.5% (w/v) |
0.5% (w/v) |
zwitterionic detergent |
| Amphipol A8-35 |
100 μl |
5% (w/v) |
|
anionic surfactant |
| Glycerol |
1 ml |
30% (w/v) |
|
cryoprotectant |
Quantifoil Holey Carbon Films
20x Quantifoil™ R 1.2/1.3 on 200 copper mesh
20x Quantifoil™ R 1.2/1.3 with ultrathin (2 nm) continuous layer of carbon on top on 200 copper mesh
20x Quantifoil™ R 2/1 on 300 copper mesh
20x Quantifoil™ R 2/1 with ultrathin (2 nm) continuous layer of carbon on top on 300 copper mesh
