Abstract
Analysis of intact multi-molecular complexes by gel electrophoresis requires the use of non-denaturing conditions. Agarose is the matrix of choice for separation of large, intact complexes because gels with larger pore sizes can be more readily prepared and manipulated compared to acrylamide. In the absence of SDS (or any other denaturant), the separation is based on both the average electrical surface charge density (σ) and the effective radius (R E). In order to separately measure σ and R E, two dimensions of electrophoresis are required. Native 1-D and 2-D agarose gel electrophoresis (AGE) have been used for analysis of bacteriophages and related particles, clathrin coated vesicles, cross-linked microtubules, multi-enzyme complexes, protein/polysaccharide vaccines, and ribosomes. 2-D AGE has proved to be especially valuable for characterization of bacteriophages and related particles because not only is information obtained about particle σ and R E, but also, the separated particles can be isolated from the agarose gel and examined by microscopy or subjected to DNA or protein analysis, as illustrated in this chapter for bacteriophage T3 proteins.
Published Version
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