A full spectrum of high-throughput protein identification and characterization approaches has been developed for protein profiling. However, the most demanding field to better understanding protein interactions known as the "interactome" is still of a perpetual need for modern proteomics. Recently developed DIGE (difference in-gel electrophoresis) system may be of potential use when studying interacting proteins. In this work we applied DIGE technique on native gel electrophoresis to study protein-protein interactions. As a proof of principle, we utilized an in vitro interaction model between p53 and HDM2 proteins. In parallel, we also showed interaction of these proteins using fluorescently labelled p53- or HDM2-immunoprecipitation pellets. Thus, we believe this study shows a good potential for investigating various interacting partners and benefits towards creation of interactome.