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Laboratory TechniquesScience

Zinc Staining Protocol

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…
July 1, 2019
Laboratory TechniquesProtocolsScience

Ponceau S Stain Protocol

Ponceau S staining is a rapid and reversible staining method used for the detection of protein bands on Western blot membranes, Polyvinylidene fluoride (PVDF), nitrocellulose, and cellulose acetate membranes. Ponceau S is a negative stain that binds to the positively charged functional groups of the protein (amino group) and the…
June 30, 2019
Laboratory TechniquesScience

Pulse Chase Analysis

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…
June 30, 2019
Laboratory TechniquesScience

Silver Staining Protocol

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…
June 30, 2019
Laboratory TechniquesProtocolsScience

Southern Blotting Protocol

Southern blotting is a hybridization technique used for deoxyribose nucleic acid (DNA). The method was named after Edward M. Southern, who developed the technique in the 1970s. The method involves the transfer of DNA fragments from an electrophoresis gel to a membrane causing immobilization of the DNA fragments and bands…
June 30, 2019
Laboratory TechniquesProtocols

Affinity Chromatography Protocol

Affinity chromatography is a versatile separation protocol that uses the biological interactions for characterization and detailed analysis of sample components. It is based on highly specific interactions between two molecules, such as the interactions between enzyme and its substrate, receptor, and ligand, or antibody and antigen. These reversible interactions are…
June 26, 2019