Determination of the Molecular Mass of Coat Proteins by Gel Electrophoresis
For determining the molecular mass of viral coat protein(s), purified virus is disrupted in denaturation buffer and the product directly applied to a polyacrylamide gel for electrophoresis. Under denaturing conditions, the viral capsid falls apart whereafter the polypeptide subunits lose their tertiary and secondary structures. Denaturation is usually with the anionic detergent SDS and 2-ME. The proteins become negatively charged and can be separated in the gel on the basis of chain length. The distance migrated is inversely proportional to the log10 of the molecular mass of the protein. Protein bands are stained and molecular masses determined by comparing migration of the viral protein(s) in the gel with that of marker proteins with known molecular mass, run in parallel lanes.
KeywordsMigration Glycerol Albumin Glycine Electrophoresis
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