two-dimensional gel electrophoresis

Two-dimensional gel electrophoresis is a powerful technique used to separate and analyze proteins. It combines two different separation methods.

First, proteins are separated based on their isoelectric point (charge) in a process called isoelectric focusing (IEF). This creates a pH gradient within a gel, causing proteins to migrate until they reach the pH where their net charge is zero.

Second, the separated proteins are then subjected to a different type of electrophoresis, often SDS-PAGE, which separates them based on their size. This two-step process allows for a much higher resolution of protein mixtures compared to single-dimension methods, making it invaluable for characterizing complex protein samples and evaluating their purity.1