Abstract
The accumulation of specific aggregation-prone proteins during aging is thought to be involved in several diseases, most notably Alzheimer’s and Parkinson’s disease as well as polyglutamine expansion disorders such as Huntington’s disease. Caenorhabditis elegans disease models with transgenic expression of fluorescently tagged aggregation-prone proteins have been used to screen for genetic modifiers of aggregation. To establish the role of modifying factors in the generation of aggregation intermediates, a method has been developed using native agarose gel electrophoresis (NAGE) that enables parallel screening of aggregation patterns of fluorescently labeled aggregation-prone proteins. Together with microscopy-based genetic screens this method can be used to identify modifiers of protein aggregation and characterize their molecular function. Although described here for analyzing aggregates in C. elegans, NAGE can be adjusted for use in other model organisms as well as for cultured cells.
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Acknowledgements
We thank Sally Hill for editing the manuscript, and Renée Sienstra and Olga Sin for critical reading of the manuscript. This project was funded by grants from the Prinses Beatrix Fonds Foundation, an NWO Meervoud grant (E.A.A.N), and a UMCG Rosalind Franklin Fellowship (E.A.A.N.).
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Holmberg, M., Nollen, E.A.A. (2013). Analyzing Modifiers of Protein Aggregation in C. elegans by Native Agarose Gel Electrophoresis. In: Hatters, D., Hannan, A. (eds) Tandem Repeats in Genes, Proteins, and Disease. Methods in Molecular Biology, vol 1017. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-438-8_14
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DOI: https://doi.org/10.1007/978-1-62703-438-8_14
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