Sensitive reverse staining of bacterial lipopolysaccharides on polyacrylamide gels by using zinc and imidazole salts.
We present a new method for visualizing bacterial lipopolysaccharides (LPS)/lipooligosaccharides (LOS) electrophoresed in sodium dodecyl sulfate-polyacrylamide gels. After electrophoresis, gels are washed in boiling water to appreciably remove remaining electrophoresis reagents, then incubated in 10 mM zinc sulfate for 15 min, and subsequently immersed in 0.2 M imidazole for 3 min. As a result, zinc salts precipitate all over the gel surface except in the zones occupied by LPS/LOS, which appear as transparent, colorless bands. Gels can be stored in distilled water for weeks without loss of the negative image. The sensitivity of this stain is similar to that of silver. We believe that zinc-imidazole may be a suitable nontoxic alternative to silver in the rapid analysis of LPS/LOS by polyacrylamide gel electrophoresis.
Hardy
E
E
Center for Genetic Engineering and Biotechnology, Havana City, Cuba.
Pupo
E
E
Castellanos-Serra
L
L
Reyes
J
J
Fernández-Patrón
C
C
D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007093:Imidazoles / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000032:analysis*; D012680:Sensitivity and Specificity; D015032:Zinc / Q000737:chemistry*
D004396:Coloring Agents; D007093:Imidazoles; D008070:Lipopolysaccharides; C029899:imidazole; D015032:Zinc