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Co-immunoprecipitation of human mitochondrial proteases AFG3L2 and paraplegin heterologously expressed in yeast cells

External protocol Created on 30 Apr 2014

Authors

Valentina Fracasso, Federico Lazzaro, and Marco Muzi-Falconi

Summary

AFG3L2 and paraplegin are cognate ATP-dependent metalloproteases that constitute the m-AAA protease complex in the inner mitochondrial membrane. Co-immunoprecipitation (CoIP) is a useful technique for the analysis of protein-protein interaction. Here, we illustrate a protocol to precipitate the complex formed by human epitope-tagged AFG3L2 and paraplegin heterologously expressed in the yeast Saccharomyces cerevisiae. The procedure employs polyclonal anti-AFG3L2 and anti-paraplegin, and monoclonal anti-HA antibodies to investigate the interaction between the two proteins. The protocol can be adapted to the analysis of other protein complexes.

Further details

The protocol was published on Protocol Exchange on 7 March 2010. To see the entire protocol, click on the source link.

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