"Reverse-staining" of biomolecules in electrophoresis gels: analytical and micropreparative applications.
Negative or reverse staining using imidazole and zinc salts for protein detection in electrophoresis gels was originally introduced in 1990. The method is based on the selective precipitation of zinc imidazolate in the gel except in the zones where proteins are located. The method was later adapted to allow high-sensitivity negative detection of nucleic acids and bacterial lipopolysaccharides. It provides a practically quantitative recovery of intact biomolecules and is a method of choice for micropreparative applications of gel electrophoresis to proteomics and similar structural studies. Zinc-mediated protein fixation in the gel is fully reversible and the eluted biomolecules are neither chemically modified nor contaminated with organic dyes. Here we present a detailed compilation of practical methods for implementing these techniques with emphasis in their analytical or micropreparative applications.
Hardy
Eugenio
E
Center for Genetic Engineering and Biotechnology, Division of Physical Chemistry, P.O. Box 6162, Havana City, Cuba.
Castellanos-Serra
Lila R
LR
D004247:DNA / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D011506:Proteins / Q000032:analysis*; D016622:Silver Staining; D013194:Staining and Labeling / Q000379:methods*
D011506:Proteins; D004247:DNA