Abstract
Mitochondria are highly dynamic organelles that are vital for eukaryotic cellular metabolism. Mitochondria account only for a small fraction of the total cell volume. As such, intact organelle isolation is a basic requirement to study mitochondrial biochemistry and physiology. However, established isolation procedures are tedious, and require a substantial amount of tissue material to start with. We addressed this matter and developed a single step purification for Arabidopsis thaliana (Arabidopsis) mitochondria. Ubiquitous expression of an 3xHA-sGFP-TOM5 fusion protein in the outer mitochondrial membrane allows affinity purification by immunocapture in a single step. We here provide a detailed step-by-step protocol for rapid immunocapture of intact and physiologically active mitochondria from Arabidopsis plants.
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Acknowledgments
This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Project-ID 267205415—SFB 1208 and under Germany’s Excellence Strategy EXC-2048/1, Project ID 390686111.
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Kuhnert, F., Weber, A.P.M. (2022). Isolation of Plant Mitochondria Using Affinity Purification. In: Van Aken, O., Rasmusson, A.G. (eds) Plant Mitochondria. Methods in Molecular Biology, vol 2363. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1653-6_3
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DOI: https://doi.org/10.1007/978-1-0716-1653-6_3
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