2DE for Proteome Analysis of Human Metaphase Chromosomes

Two-dimensional electrophoresis (2DE) is one of the most effective methods for the reliable separation of proteins in a single gel. In our proteome analyses of human chromosomes, we used two types of 2DE: two-dimensional isoelectric focusing SDS–polyacrylamide gel electrophoresis (2D IEF/SDS–PAGE) and radical-free and highly reduced two-dimensional polyacrylamide gel electrophoresis (RFHR 2D PAGE) together with one-dimensional SDS–polyacrylamide gel electrophoresis (1DE). Experimental details of these gel electrophoresis procedures that have been shown to be effective for human proteome analyses are described in detail.