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PAGE analysis of lipopolysaccharide (LPS) and K-antigens (KPS)

NT Nicolás Toro
PV Pablo J. Villadas
MM María Dolores Molina-Sánchez
PN Pilar Navarro-Gómez
JV José M. Vinardell
LC Lidia Cuesta-Berrio
MR Miguel A. Rodríguez-Carvajal
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LPS extraction from bacterial cultures grown on solid TY medium, separation on SDS-PAGE gels and silver staining were performed as previously described39. K-antigen capsular polysaccharides (KPS) were extracted from bacterial cultures grown on solid TY medium, as previously described40. Samples were analyzed by PAGE, as previously described41, except that absolute ethanol was added to the running buffer, and the running and stacking gels, at a final concentration of 10% (v/v) in all cases. The acrylamide concentration of the running gel was 18% (w/v), and the acrylamide/N,N′-methylene bisacrylamide ratio was 29:1. Gels were fixed with Alcian Blue (0.5% in 2% acetic acid) and silver-stained.

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