Coomassie brilliant blue (CBB) stain is a widely used method for routine visualization of proteins separated on polyacrylamide gels. It is an organic dye that makes complexes with basic amino acids, such as lysine, histidine, tyrosine, and arginine. The stain transfers an overall negative charge to the proteins allowing their…

Protein separation and identification is critical for proteome analysis and requires high resolution and powerful protein characterization after gel electrophoresis. To identify the proteins in the gel, colorimetric and fluorescence staining techniques are used. The stains include anionic dyes (Coomassie brilliant blue), metal cations (imidazole-zinc), silver stain, fluorescent dyes, and…

The reversible Zinc stain is a widely used staining method for the detection of proteins fractionated by polyacrylamide gels or SDS denatured gels. The Zinc reverse staining exploits the ability of biopolymers to bind Zn2+ and that of imidazole to react with unbound Zn2+ to produce insoluble zinc-imidazolate (ZnIm2). This…

The pulse-chase analysis is a powerful technique to study the synthesis, processing, and transport of proteins. The pulse-chase analysis is widely used in biochemistry and molecular biology for the examination of the cellular process by exposing the cells to a labeled compound (pulse). The labeled compound is introduced into the…

Silver staining is a powerful technique for protein identification in gels as silver binds to chemical sidechains of the amino acids, including the carboxyl and sulfhydryl groups. It was introduced in 1972 and later adapted for protein separation from the polyacrylamide gel electrophoresis. The nucleation sites in proteins promote the…