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Purification of tubulin from limited volumes of cultured cells.

Anne Fourest-Lieuvin 1, *
* Corresponding author
1 ANTE-INSERM U836, équipe 4, Muscles et pathologies
Organisation Fonctionnelle du Cytosquelette
Abstract : A method was designed to purify tubulin from limited volumes of cultured cells, which can be performed in less than 4 h. The method is based on the preservation of intact microtubule arrays during cell lysis in a large volume of buffer, followed by disassembly of microtubules in a small volume of cold buffer. This allows a good enrichment in tubulin, which is then purified by one cycle of polymerisation/depolymerisation and a cation exchange chromatography. Such a procedure has been employed successfully on suspension-cultured and on adherent HeLa cells. Tubulin obtained was 90% pure, assembly-competent and composed of alpha/beta I and alpha/beta IV isotypes. Microtubules made with this tubulin displayed specific properties such as resistance to dilution, maybe related to their specific dynamic behaviour.
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Anne Fourest-Lieuvin. Purification of tubulin from limited volumes of cultured cells.. Protein Expression and Purification, Elsevier, 2006, 45 (1), pp.183-90. ⟨10.1016/j.pep.2005.05.011⟩. ⟨inserm-00380060⟩

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