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Visualizing Proteins and Macromolecular Complexes by Negative Stain EM

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

Article DOI: 10.3791/3227-v 08:02 min December 22nd, 2011
December 22nd, 2011



Visualizing protein samples by negative stain electron microscopy (EM) has become a popular structural analysis method. It is useful for quantitative structural analysis, such as calculating a 3D reconstruction of the molecules being studied, and also for qualitative examination of the quality of protein preparations. In this article we present detailed protocols for preparing the EM grids, staining the sample and visualizing the sample in an electron microscope. Novice users can follow these protocols easily and to utilize negative stain EM as a routine assay, in addition to other biochemical assays, for evaluating their protein samples.


Visualizing Proteins Macromolecular Complexes Negative Stain EM Grid Preparation Image Acquisition Single Particle Electron Microscopy Structural Biology Protein Structures CryoEM Sample Preparation Method Dried Heavy Metal Salt Specimen Contrast Negative Stain EM Three-dimensional Structure Purified Proteins Protein Complexes Homogeneity/heterogeneity Large Assemblies Quality Evaluation EM Protocol Carbon Coated Grids Electron Microscope
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