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Laboratory TechniquesScience
July 3, 2019

Protein Staining Methods

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…
Laboratory TechniquesScience
July 1, 2019

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…
Laboratory TechniquesProtocolsScience
June 30, 2019

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…
Laboratory TechniquesScience
June 30, 2019

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…
Laboratory TechniquesScience
June 30, 2019

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…
Laboratory TechniquesProtocolsScience
June 30, 2019

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…
Laboratory TechniquesProtocols
June 26, 2019

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…
Laboratory TechniquesProtocols
June 26, 2019

Fast protein liquid chromatography (FPLC) Protocol

Fast protein liquid chromatography (FPLC) is a type of liquid chromatography that provides high resolution by small-diameter stationary phases for protein characterization and separation. The technique features high loading capacity, biocompatible buffer systems, fast flow rates, and stationary phases for common chromatography modes (e.g., gel filtration, ion exchange, reversed phase,…