Protein-DNA Binding Assay (AlphaScreen Technology).

Identification of specific DNA binding sites of transcription factors is important in understanding their functions. Recent techniques allow us to investigate genome-wide binding positions by chromatin immunoprecipitation combined with high-throughput sequencing. However, to further explore the binding motifs of transcription factors, in-depth biochemical analysis is required. Here, we describe an efficient protocol of protein-DNA interactions based on a combination of our transcription/translation system and AlphaScreen technology. The transcription/translation system supports an efficient and quick way of protein synthesis by alleviating cumbersome cloning steps. In addition, AlphaScreen system provides a highly sensitive, quick, and easy handling platform to investigate the protein-DNA interactions . Thus, our method largely contributes to comprehensive analysis of the biochemical properties of transcription factors.