Negative detection of biomolecules separated in polyacrylamide electrophoresis gels.
Here we describe the protocols for negative or reverse detection of proteins, nucleic acids and lipopolysaccharides separated in polyacrylamide electrophoresis gels. These protocols are based on the selective synthesis and precipitation of a white imidazole-zinc complex in the gel, which is absent from those zones where biomolecules are located. These methods are highly sensitive (1-10 ng of biomolecules per band), very cheap as they use inexpensive, common laboratory reagents (imidazole and a Zn II salt), rapid (less than 20 min after gel washing), robust and simple (two steps). Reverse-stained biomolecules are reversibly fixed in the gel. After brief incubation in a zinc chelating agent, biomolecules can be recovered from the gel with the same efficiency as from unstained gels. In consequence, they are procedures of choice for micropreparative applications. References covering typical applications are included.
Castellanos-Serra
Lila
L
Division of Physical Chemistry, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Hardy
Eugenio
E
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007093:Imidazoles; D008070:Lipopolysaccharides / Q000032:analysis*; D009696:Nucleic Acids / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D017967:Zinc Compounds
D007093:Imidazoles; D008070:Lipopolysaccharides; D009696:Nucleic Acids; D011506:Proteins; D017967:Zinc Compounds; C029899:imidazole