Detection of biomolecules in electrophoresis gels with salts of imidazole and zinc II: A decade of research

The proven ability of gel electrophoresis to simultaneously resolve, in a s ingle experiment, many components from complex biological samples, has dete rmined its preference over a variety of well-established chromatographic me thods, Therefore, procedures placed at the interface between gel separation and microanalysis have earned increasing significance with respect to the overall success of the microanalytical strategy. The first of these procedu res is the detection technique. The most important requirement for compatib ility with further analysis or bioapplications is that the staining method does not compromise the chemical integrity and the biological properties of micropurified biomolecules. Procedures for negative detection of proteins with metal salts that have been proven to comply with this condition have b een known for about 15 years. Only recently have these procedures been exte nded to the field of nucleic acids and lipopolysaccharides. The focus of th is review is to chronicle the development and current status of the negativ e or reverse stain procedure based on the in-gel reaction of imidazole with zinc salts and its applications for the micropurification and analysis of unmodified proteins, nucleic acids and bacterial lipopolysaccharides. We hi ghlight the common aspects in the detection of the three types of biomolecu les, and their applications to structural and biological analyses. Emphasis is given on the mechanism underlying imidazole-zinc staining, as it contri butes to a deeper understanding of a general detection mechanism with metal salts. Finally, we discuss the latest applications of the techniques in pr oteomics and their possible impact on the characterization of gel-separated single components from complex lipopolysaccharides.