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

✍ Scribed by Thomas J. Inzana; Michael A. Apicella

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 79 KB
• Volume: 20
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/(sici)1522-2683(19990301)20:3<462::aid-elps462>3.0.co;2-n

✦ Synopsis

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.