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Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition
Published on: December 22, 2011
Negative-Stain Immunoelectron-Microscopic Analysis of Small Macromolecules of Immunologic Significance
Roux1
1Department of Biological Science and Structural Biology Program, Florida State University, Tallahassee, Florida, 32306
Abstract:
We have demonstrated that unlabeled antibodies or their antigen-reactive fragments, Fab and F(ab')2, can be readily visualized by negative-stain immunoelectron microscopy and can be quite useful as probes for detailed epitope mapping and structural analysis of relatively small macromolecules. In addition, information on segmental flexibility of the target molecules can be deduced as can information on the geometry and kinetics of immune complex formation in solution. The various target molecules used to illustrate these points include IgG, IgM, C-reactive protein, and complement receptor 1.
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