Structural Analysis of Proteins by Transmission Electron Microscopy

  • Different orientations of IgG antibodies on a NS-TEM grid; 40 class averages are shown (30 particle projections per average).

  • Cholesteryl ester transfer protein (CETP) labeled by Fab fragments from three different monoclonal antibodies.

About-+

In this project we focus on negative-stain transmission electron microscopy (NS-TEM) and the localization of specific domains/components of a protein complex by labeling techniques, e.g., by the use of antibodies and fragments thereof (Fabs, which have a mass of just 50 kDa).

More detail+-

The structure, shape, 3D conformation, homogeneity and aggregation state of the macromolecules in a protein sample can be evaluated and assessed by negative-stain transmission electron microscopy (NS-TEM) using only minute amounts of sample (just a few microliters of the protein solution).

The single molecule imaging approach can be used to document and extend the findings of biochemical experiments performed on bulk solutions (e.g., size exclusion chromatography or analytical ultracentrifugation), and to obtain initial structural information (20Å resolution).

Besides being a quick and efficient method to assess the quality of protein samples, NS-TEM allows the proteins/protein complexes present to be characterized further by the use of labels, such as nanogold complexes, quantum dots, antibodies or Fabs.  

Related Publications+-

Related Publications