Protocol Protocol for separation of the nuclear and the cytoplasmic fractions of Xenopus laevis embryonic cells for studying protein shuttling

This protocol for the separation of nuclear and cytoplasmic fractions of cells of Xenopus laevis embryos was developed to study changes in the intracellular localization of the Zyxin and Ybx1 proteins, which are capable of changing local-ization in response to certain stimuli. Western blot analysis allows the quantifica-tion of changes in the distribution of these proteins between the cytoplasm and nucleus, whereas the posttranslational modifications specific to each compart-ment can be identified by changes in electrophoretic mobility. For complete details on the use and execution of this protocol, please refer to Parshina et al. (2020).

Automated summary

This protocol is developed for the separation of nuclear and cytoplasmic fractions of Xenopus laevis embryo cells to study changes in intracellular localization of Zyxin and Ybx1 proteins. Western blot analysis allows quantification of protein distribution changes between cytoplasm and nucleus, while posttranslational modifications specific to each compartment can be identified by changes in electrophoretic mobility.