Electrophoretic mobility shift assays (EMSAs) for in vitro detection of protein-nucleic acid interactions

Abstract

Summary: Protein-nucleic acid interactions drive some of the most important physiological events in cells. Here, we present a protocol for detecting protein-DNA or protein-RNA interactions in vitro. We describe steps for labeling nucleic acid species and electrophoretic mobility shift assays (EMSAs). This protocol can be used to confirm suspected in vivo interactions using recombinantly expressed/purified proteins of interest and a nucleic acid substrate. It can further be used to investigate mutations that can disrupt interaction or compensatory mutations that restore it.For complete details on the use and execution of this protocol, please refer to Mansouri-Noori et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Molecular Biology
• Gene Expression
• Molecular/Chemical Probes
• Protein Biochemistry