Purification of Affinity Tag-free Recombinant Tubulin from Insect Cells

被引:7
|
作者
Ti, Shih-Chieh [1 ,2 ]
Wieczorek, Michal [1 ]
Kapoor, Tarun M. [1 ]
机构
[1] Rockefeller Univ, Lab Chem & Cell Biol, 1230 York Ave, New York, NY 10065 USA
[2] Univ Hong Kong, LKS Fac Med, Sch Biomed Sci, Hong Kong, Peoples R China
来源
STAR PROTOCOLS | 2020年 / 1卷 / 01期
关键词
ONE-STEP PURIFICATION; DYNAMIC INSTABILITY; INDIVIDUAL MICROTUBULES; MICROSCOPY;
D O I
10.1016/j.xpro.2019.100011
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
a/8-tubulin heterodimers, which can harbor diverse isotypes and post -translational modifications, polymerize into microtubules that are fundamental to many cellular processes. Due to long-standing challenges in generating recombinant tubulin, however, it has been difficult to examine the properties of specific tubulin isotypes. Here, we provide a protocol for purifying milligrams of affinity tag-free, isotypically pure recombinant tubulin. Our method can be applicable to any tubulin of interest, opening the door to dissecting how tubulin diversity regulates microtubule function.For complete details on the use and execution of this protocol, please see Ti et al. (2018).
引用
收藏
页数:25
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