Fluorescein-Based Electrophoretic Mobility Shift Assay

Shachinthaka D Dissanayaka Mudiyanselage; Ying Wang

doi:10.1007/978-1-0716-1464-8_12

Fluorescein-Based Electrophoretic Mobility Shift Assay

Methods Mol Biol. 2022:2316:133-140. doi: 10.1007/978-1-0716-1464-8_12.

Authors

Shachinthaka D Dissanayaka Mudiyanselage¹, Ying Wang²

Affiliations

¹ Department of Biological Sciences, Mississippi State University, Mississippi State, MS, USA.
² Department of Biological Sciences, Mississippi State University, Mississippi State, MS, USA. wang@biology.msstate.edu.

PMID: 34845691
DOI: 10.1007/978-1-0716-1464-8_12

Abstract

RNA-protein complexes are functionally important in biology. Electrophoretic mobility shift assays (EMSA) have been widely used to study the molecular basis of protein-RNA interactions. Previous methods for EMSA mostly relied on radioactive RNA substrates, raising health and environmental concerns. Here, we describe a method based on fluorescein-labeled RNA for EMSA. In addition, we simplified the protocol to efficiently purify RNA-binding proteins from bacterial expression systems.

Keywords: EMSA; Fluorescein; Viroid.

MeSH terms

Electrophoretic Mobility Shift Assay*
Fluorescein
Protein Binding
RNA / metabolism
RNA-Binding Proteins

Substances

RNA-Binding Proteins
RNA
Fluorescein