Variations on a theme: Changes to electrophoretic separations that can make a difference

TL;DR

This paper reviews various modifications to electrophoretic separation protocols, highlighting how adjustments in gel, buffer, and detergents can enhance protein separation in proteomics.

Contribution

It provides a comprehensive overview of key variations in electrophoretic methods, aiding researchers in optimizing protein separation techniques.

Findings

Identifies critical parameters affecting electrophoretic separation quality.

Summarizes modifications that improve resolution and reproducibility.

Highlights potential advantages of alternative electrophoretic approaches.

Abstract

Electrophoretic separations of proteins are widely used in proteomic analyses, and rely heavily on SDS electrophoresis. This mode of separation is almost exclusively used when a single dimension separation is performed, and generally represents the second dimension of two-dimensional separations. Electrophoretic separations for proteomics use robust, well-established protocols. However, many variations in almost all possible parameters have been described in the literature over the years, and they may bring a decisive advantage when the limits of the classical protocols are reached. The purpose of this article is to review the most important of these variations, so that the readers can be aware of how they can improve or tune protein separations according to their needs. The chemical variations reviewed in this paper encompass gel structure, buffer systems and detergents for SDS electrophoresis, two-dimensional electrophoresis based on isoelectric focusing and two-dimensional electrophoresis based on cationic zone electrophoresis.