Affinity purification of label-free tubulins from xenopus egg extracts

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dc.contributor.author Reusch, Sebastian
dc.contributor.author Biswas, Abin
dc.contributor.author Hirst, William G.
dc.contributor.author Reber, Simone
dc.date.accessioned 2021-03-16T20:55:01Z
dc.date.available 2021-03-16T20:55:01Z
dc.date.issued 2020-12-18
dc.description © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Reusch, S., Biswas, A., Hirst, W. G., & Reber, S. Affinity purification of label-free tubulins from xenopus egg extracts. STAR Protocols, 1(3), (2020): 100151, doi:10.1016/j.xpro.2020.100151. en_US
dc.description.abstract Cytoplasmic extracts from unfertilized Xenopus eggs have made important contributions to our understanding of microtubule dynamics, spindle assembly, and scaling. Until recently, these in vitro studies relied on the use of heterologous tubulin. This protocol allows for the purification of physiologically relevant Xenopus tubulins in milligram yield, which are a complex mixture of isoforms with various post-translational modifications. The protocol is applicable to any cell or tissue of interest. For complete details on the use and execution of this protocol, please refer to Hirst et al. (2020). en_US
dc.description.sponsorship This article was prompted by our stay at the Marine Biological Laboratory (MBL), Woods Hole, MA, in the summer of 2016 funded by the Princeton-Humboldt Strategic Partnership Grant together with the lab of Sabine Petry (Princeton University). We are grateful to the National Xenopus Resource (NXR) for supplying frogs. For mass spectrometry, we would like to acknowledge the assistance of Benno Kuropka and Chris Weise from the Core Facility BioSupraMol supported by the Deutsche Forschungsgemeinschaft (DFG). We thank the Protein Expression Purification and Characterization (PEPC) facility at the MPI-CBG; in particular, we thank Aliona Bogdanova and Barbara Borgonovo. We thank all former and current members of the Reber lab for discussions and helpful advice, in particular Christoph Hentschel and Soma Zsoter for technical assistance. S.R. acknowledges funding from the IRI Life Sciences (Humboldt-Universität zu Berlin, Excellence Initiative/DFG). W.H. was supported by the Alliance Berlin Canberra co-funded by a grant from the Deutsche Forschungsgemeinschaft (DFG) for the International Research Training Group (IRTG) 2290 and the Australian National University. en_US
dc.identifier.citation Reusch, S., Biswas, A., Hirst, W. G., & Reber, S. (2020). Affinity purification of label-free tubulins from xenopus egg extracts. STAR Protocols, 1(3), 100151. en_US
dc.identifier.doi 10.1016/j.xpro.2020.100151
dc.identifier.uri https://hdl.handle.net/1912/26818
dc.publisher Cell Press en_US
dc.relation.uri https://doi.org/10.1016/j.xpro.2020.100151
dc.rights Attribution-NonCommercial-NoDerivatives 4.0 International *
dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/ *
dc.title Affinity purification of label-free tubulins from xenopus egg extracts en_US
dc.type Article en_US
dspace.entity.type Publication
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relation.isAuthorOfPublication.latestForDiscovery 19c7af05-27e8-4682-9d95-7d94f43efef0
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