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剥离印迹技术：一种冷冻电镜样品制备方法，用于从多层或浓缩生物样品中分离单层

JoVE Journal
Bioquímica

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JoVE Journal Bioquímica

The Peel-Blot Technique: A Cryo-EM Sample Preparation Method to Separate Single Layers From Multi-Layered or Concentrated Biological Samples

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剥离印迹技术：一种冷冻电镜样品制备方法，用于从多层或浓缩生物样品中分离单层

DOI:

10.3791/64099-v

•

07:27 min

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June 29, 2022

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Matthew C. Johnson*^1,2, Arshay J. Grant*¹, Ingeborg Schmidt-Krey³

¹School of Biological Sciences,Georgia Institute of Technology, ²Department of Structural Biology,Genentech, ³School of Chemistry and Biochemistry,Georgia Institute of Technology

Capítulos

00:04Introduction
00:33Preparation Steps for the Peel‐Blot Technique
01:57Placing the Carbon Film onto the Grid
03:07Separation of Multi-Layered 2D Crystals into Single Layer
04:55Results: 2D Crystals of Human Leukotriene C4 Synthase and Liposomes Subjected to the Peel‐Blot Method, the Peel-Blot Footprint, and Blotting an EM Grid Prepared by Back-Injection on a Submicron Filter Paper
06:42Conclusion

Summary

Tadução automática

剥离-印迹技术是一种冷冻电镜网格制备方法，可将多层和浓缩的生物样品分离成单层，以减小厚度、提高样品浓度并促进图像处理。

Tags

Keywords: Peel-Blot Technique Cryo-EM Sample Preparation Multi-Layered Samples Concentrated Samples Single Layers Thickness Reduction Concentration Increase Image Processing Filter Paper Paraffin Film Submicron Filter Paper Anti-Capillary Forceps 600 Mesh Grid Trehalose Solution Mica Carbon Film Grid Orientation

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