Abstract
FAT10 plays a role in many cellular processes. Nevertheless, only one substrate could be identified so far to which FAT10 becomes covalently attached via a nonreducible isopeptide bond. The identification of additional substrates as well as interaction partners is therefore of great interest. Due to the absence of potent anti-FAT10 antibodies, the detection and identification of interaction partners was hindered so far. We have recently described the generation of a novel monoclonal FAT10 antibody that is suitable for immunoprecipitation of endogenous FAT10 and describe in this chapter the detection of endogenous as well as of His-3×FLAG-tagged FAT10 interacting proteins and conjugates by immunoprecipitation using either this novel anti-FAT10 antibody 4FI or an anti-FLAG affinity gel.
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Acknowledgments
This work was supported by grants of the German Research Foundation (DFG) GR 1517/9-1 and GR 1517/10-1, the Thurgauische Stiftung für Wissenschaft und Forschung, and the Swiss State Secretariat for Education and Research.
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Aichem, A., Groettrup, M. (2012). Detection and Analysis of FAT10 Modification. In: Dohmen, R., Scheffner, M. (eds) Ubiquitin Family Modifiers and the Proteasome. Methods in Molecular Biology, vol 832. Humana Press. https://doi.org/10.1007/978-1-61779-474-2_7
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DOI: https://doi.org/10.1007/978-1-61779-474-2_7
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