ABSTRACT
This chapter describes how to prepare sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels and their use for separating and resolving bacterial proteins. Although there exist a range of procedures and types of apparatus used for SDS-PAGE, the chapter focuses on the method described by U. K. Laemmli and used in association with a Shandon vertical slab gel apparatus. The SDS-PAGE gel preparation procedure relates to the Shandon Model 200 slab electrophoresis system. Conventional electrophoresis systems give very good separation of bacterial components due to the length of the separation gel. When electrophoresis has been completed, pour Coomassie blue stain or silver stain fixing solution into a clean plastic box of a size sufficient for free movement of gels during agitation. A gel electrophoresis system comprises a set of gel plates, sample well-forming combs, gel electrophoresis tank and clips for clamping gel plates together.
