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Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example
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Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example

Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example

DOI:
10.3791/52230-v

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12:44 min

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December 03, 2014

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, ,
1Lady Davis Institute for Medical Research,Jewish General Hospital, 2Department of Medicine,McGill University

Chapters

  • 00:05Title
  • 01:28Preparation of Protein Extracts from Cultured Cells
  • 03:05Preparation of Protein Extracts from Mouse Liver and Spleen
  • 04:56Synthesis of Radiolabeled IRE-containing RNA Probe
  • 05:55Purification of Radiolabeled IRE-containing RNA Probe
  • 07:48Electrophoretic Mobility Shift Assay (EMSA)
  • 10:27Results: The Iron-dependence of IRE/IRP Interactions
  • 12:17Conclusion

Summary

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Here we present a protocol to analyze RNA/protein interactions. The electrophoretic mobility shift assay (EMSA) is based on the differential migration of RNA/protein complexes and free RNA during native gel electrophoresis. By using a radiolabeled RNA probe, RNA/protein complexes can be visualized by autoradiography.

Tags

Keywords: RNA-protein InteractionsIron Regulatory ProteinsIRP1IRP2Iron Responsive ElementsIREUTRsEMSAGel RetardationBandshift AssayRNA-binding ProteinsProtein-RNA ComplexesPost-transcriptional Regulation

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