从培养的人细胞中分离有丝分裂中心体。
Isolation of Mitotic Centrosomes from Cultured Human Cells.
机构信息
Department of Physiological Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Division of Molecular Oncology, Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
出版信息
Methods Mol Biol. 2025;2872:233-244. doi: 10.1007/978-1-0716-4224-5_16.
The centrosome plays a crucial role in facilitating mitotic spindle assembly through its microtubule organizing capacities. Analyzing the composition, structure, and functions of mitotic centrosomes is essential for understanding the mechanisms underlying cell division and centrosome-associated diseases. Isolating centrosomes is an effective method to gain comprehensive information about them while minimizing interference from other cellular components. In this chapter, we describe a protocol for isolating mitotic centrosomes from cultured human cells. This protocol includes cell synchronization and centrosome isolation through ultracentrifugation with a sucrose gradient. We also describe a method for conducting a microtubule nucleation assay to assess the functionality of isolated centrosomes.
中心体通过其微管组织能力在促进有丝分裂纺锤体组装中起着至关重要的作用。分析有丝分裂中心体的组成、结构和功能对于理解细胞分裂和中心体相关疾病的机制至关重要。分离中心体是获得有关它们的全面信息而最大限度地减少来自其他细胞成分干扰的有效方法。在本章中，我们描述了一种从培养的人细胞中分离有丝分裂中心体的方案。该方案包括通过蔗糖梯度超速离心进行细胞同步化和中心体分离。我们还描述了一种进行微管核形成测定的方法，以评估分离的中心体的功能。
Zotero 插件