Affinity Purification of Label-free Tubulins from Xenopus Egg Extracts

STAR Protoc. 2020 Oct 31;1(3):100151. doi: 10.1016/j.xpro.2020.100151. eCollection 2020 Dec 18.

Abstract

Cytoplasmic extracts from unfertilized Xenopus eggs have made important contributions to our understanding of microtubule dynamics, spindle assembly, and scaling. Until recently, these in vitro studies relied on the use of heterologous tubulin. This protocol allows for the purification of physiologically relevant Xenopus 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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Extracts / chemistry*
  • Chromatography, Affinity / methods*
  • Female
  • Ovum / cytology*
  • Protein Domains
  • Staining and Labeling*
  • Tubulin / chemistry
  • Tubulin / isolation & purification*
  • Xenopus / metabolism*

Substances

  • Cell Extracts
  • Tubulin