IRI Life Sciences, Humboldt-Universität zu Berlin, 10115 Berlin, Germany.
Marine Biological Laboratory, Woods Hole, MA 02543, USA.
STAR Protoc. 2020 Oct 31;1(3):100151. doi: 10.1016/j.xpro.2020.100151. eCollection 2020 Dec 18.
Cytoplasmic extracts from unfertilized eggs have made important contributions to our understanding of microtubule dynamics, spindle assembly, and scaling. Until recently, these studies relied on the use of heterologous tubulin. This protocol allows for the purification of physiologically relevant 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).
未受精卵的细胞质提取物对我们理解微管动力学、纺锤体组装和比例有重要贡献。直到最近，这些研究还依赖于异源微管蛋白的使用。本方案允许以毫克产量纯化生理相关的微管蛋白，这些微管蛋白是具有各种翻译后修饰的同工型的复杂混合物。该方案适用于任何感兴趣的细胞或组织。有关该方案使用和执行的完整详细信息，请参见 Hirst 等人（2020 年）。
