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

53.1K Views

12:44 min

December 3rd, 2014

December 3rd, 2014

53,070 Views

0:05

Title

1:28

Preparation of Protein Extracts from Cultured Cells

3:05

Preparation of Protein Extracts from Mouse Liver and Spleen

4:56

Synthesis of Radiolabeled IRE-containing RNA Probe

5:55

Purification of Radiolabeled IRE-containing RNA Probe

7:48

Electrophoretic Mobility Shift Assay (EMSA)

10:27

Results: The Iron-dependence of IRE/IRP Interactions

12:17

Conclusion

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.

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