Electrophoretic mobility shift assays with GFP-tagged proteins (GFP-EMSA)
Other Publication ResearchOnline@JCUThe electrophoretic mobility shift assay (EMSA) is commonly used for the study of nucleic acid-binding proteins. The technique can be used to demonstrate that a protein is binding to RNA or DNA through visualization of a shift in electrophoretic mobility of the nucleic acid band. A major disadvantage of the EMSA is that it does not always provide an absolute certitude that the band shift is due to the protein under scrutiny, as contaminants in the sample could also cause the band shift. Here we describe a variation of the standard EMSA allowing to visualize with added certitude, the co-localized band shifts of a GFP-tagged protein binding to its cognate nucleic acid target sequence stained with an intercalator, such as GelRed. Herein, we present an illustrative protocol of this useful technique called GFP-EMSA along with specific notes on its advantages and limitations.
Methods in Molecular Biology
Targeting Enzymes for Pharmaceutical Development
2089
1064-3745
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8
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Springer
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New York, NY, USA
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10.1007/978-1-0716-0163-1_10