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Electrophoresis. 1999 Mar;20(3):462-5.

Use of a bilayer stacking gel to improve resolution of lipopolysaccharides and lipooligosaccharides in polyacrylamide gels.

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1
Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg 24061, USA. tinzana@vt.edu

Abstract

Lipopolysaccharide (LPS) and lipooligosaccharide (LOS) are important antigenic and integral components of the outer membrane of Gram-negative bacteria. Alteration or heterogeneity of LPS/LOS structure is most often assessed by alteration of electrophoretic band profiles using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In order to discern minor differences in the electrophoretic profile of closely spaced bands, particularly the low molecular weight bands of LOS, optimum resolution is required. Unfortunately, many publications of LPS/LOS in polyacrylamide gels show a diffuse, smeared pattern without discernible bands. We report here a formulation for polyacrylamide gels that reproducibly yields LPS/LOS bands with sharp resolution. A key feature of this formulation is the use of a separate comb gel containing electrode buffer layered on top of the conventional stacking gel.

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