Protein-DNA Binding Assay

Clontech’s new Protein-DNA Binding Assay provides a safe, fast, and sensitive alternative to traditional electromobility shift assays (EMSA) for detection and quantitative characterization of protein-DNA interactions.

This assay eliminates the need for radioactive labeling and gel electrophoresis, since it utilizes sensitive, quantitative, chemiluminescent ProLabel™ detection, and can be performed in a 96-well plate. A small (~6 kDa) ProLabel tag is fused to your protein of interest to generate a fusion protein that is capable of producing a strong chemiluminescent signal via an enzyme complementation assay. The ProLabel tag allows direct detection of specific binding between your protein of interest and a biotinylated dsDNA oligonucleotide. The biotin moiety on the oligonucleotide permits the resulting protein-DNA complex to be captured on the 96-well plate, which is coated with streptavidin. Since the ProLabel fusion protein is expressed in mammalian cells, it can acquire the endogenous posttranslational modifications that are often needed to capture biologically relevant protein-DNA interactions. Moreover, the Protein-DNA Binding Assay utilizes whole cell lysates, eliminating the need for nuclear extraction.