Two-dimensional gel electrophoresis in proteomics: A tutorial

TL;DR

This tutorial explains the history, methodology, advantages, limitations, and applications of two-dimensional gel electrophoresis in proteomics, highlighting its continued relevance despite newer techniques.

Contribution

It provides a comprehensive guide to the technique's steps, constraints, and optimal applications, with detailed literature examples, aiding researchers in proteomics.

Findings

Highlights the unique ability to separate complete proteins.

Discusses interfacing with immunoblotting techniques.

Analyzes the technique's limitations and advantages.

Abstract

Two-dimensional electrophoresis of proteins has preceded, and accompanied, the birth of proteomics. Although it is no longer the only experimental scheme used in modern proteomics, it still has distinct features and advantages. The purpose of this tutorial paper is to guide the reader through the history of the field, then through the main steps of the process, from sample preparation to in-gel detection of proteins, commenting the constraints and caveats of the technique. Then the limitations and positive features of two-dimensional electrophoresis are discussed (e.g. its unique ability to separate complete proteins and its easy interfacing with immunoblotting techniques), so that the optimal type of applications of this technique in current and future proteomics can be perceived. This is illustrated by a detailed example taken from the literature and commented in detail. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP 2).