Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan.
Faculty of Science, Hokkaido University, Sapporo, Japan.
Methods Mol Biol. 2022;2430:3-16. doi: 10.1007/978-1-0716-1983-4_1.
Filamentous microtubules, polymers of the heterodimeric protein tubulins play one of the major roles in the emergent nano-biotechnological devices. To develop the feature of those devices, it is important to understand the function of microtubule in in vitro, hence, the availability of purified αβ-tubulin is required. Additionally, fluorescently labeled tubulin has become a powerful approach for extensively studying the dynamics of these components. In this chapter, the process of purifying the heterodimeric αβ-tubulin from porcine brain will be described, as well as the process of labeling of the purified tubulin with fluorescence dye.
丝状微管是异源二聚体蛋白微管蛋白的聚合物，在新兴的纳米生物技术设备中发挥着主要作用之一。为了开发这些设备的功能，了解微管在体外的功能非常重要，因此需要获得纯化的αβ-微管蛋白。此外，荧光标记的微管蛋白已成为广泛研究这些成分动力学的有力方法。本章将描述从猪脑中纯化异源二聚体αβ-微管蛋白的过程，以及用荧光染料标记纯化微管蛋白的过程。
