Affinity Purification of Label-free Tubulins from Xenopus Egg Extracts
Sebastian Reusch, Abin Biswas, William G. Hirst, Simone Reber
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).
Topics & Concepts
XenopusGene isoformMicrotubuleTubulinComputational biologyHeterologousBiologyCytoplasmCell biologyChemistryMolecular biologyBiochemistryGeneMicrotubule and mitosis dynamicsProtist diversity and phylogenyPhotosynthetic Processes and Mechanisms