Ultrapure water improves results in 2D gel electrophoresis Water Makes the Difference

Are there differences in quality between stored and freshly prepared ultrapure water? This article presents the results for the use of ultrapure water in gel electrophoresis and provides recommendations for the production of ultrapure water.

Two-dimensional gel electrophoresis, called 2D PAGE for short, was developed by Klose and O’Farrell independently of each other in 1975 [1, 2]. It is used to separate protein mixtures into individual proteins and today has become an indispensable method in protein analysis (proteomics).

This 2D procedure consists of two separation principles, known as dimensions (see Fig. 1). While in the first dimension, ampholytes or immobilizing pH gradients (IPGs) are used to separate proteins according to their isoelectric points, in the subsequently second dimension, proteins are separated according to their molecular weights in SDS PAGE. Afterwards, the separated proteins are visualized using different staining methods, (Coomassie blue, silver staining, etc.) and employed to perform subsequent downstream analysis, as needed.