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Label-Free Electrophoretic Mobility Shift Assay (EMSA) for Measuring Dissociation Constants of Protein-RNA Complexes.

Curr Protoc Nucleic Acid Chem. 2019-03; 
SeoMinguk,LeiLi,EgliMa
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PCR Cloning and Subcloning An expression system for human Ago2 PAZ (amino acids 227 to 371; Fig. 1B) was generated by gene synthesis (GenScript Inc.) and subcloning into the pHD116 plasmid. Get A Quote

摘要

The electrophoretic mobility shift assay (EMSA) is a well-established method to detect formation of complexes between proteins and nucleic acids and to determine, among other parameters, equilibrium constants for the interaction. Mixtures of protein and nucleic acid solutions of various ratios are analyzed via polyacrylamide gel electrophoresis (PAGE) under native conditions. In general, protein-nucleic acid complexes will migrate more slowly than the free nucleic acid. From the distributions of the nucleic acid components in the observed bands in individual gel lanes, quantitative parameters such as the dissociation constant (K ) of the interaction can be measured. This article describes a simple and r... More

关键词

Ago2,RNA,electrophoresis,equilibrium constant,protein-nucleic acid interac