PURPOSE: The aim of this article is to describe the technical development in proteomics during the last two decades with the focus on its use in radiation biology. It is written from a subjective point of view and aims not to be a scientific review of the subject. CONCLUSION: Proteomics is a fast developing technique and it has already contributed greatly to our understanding of biological mechanisms following radiation exposure. Novel proteomics approaches can be used in adequately designed cellular and animal experiments and above all in big clinical trials to investigate effects of ionizing radiation in the future.