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

• Published in: STAR protocols Vol. 2; no. 2; p. 100449
• Main Authors: Martynova, Natalia Y., Parshina, Elena A., Zaraisky, Andrey G.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.06.2021; Elsevier
• Subjects: Cell Biology; Cell separation/fractionation; Model Organisms; Molecular Biology; Protein Biochemistry; Protocol; Cell separation/fractionation; Model Organisms; Protein Biochemistry; Molecular Biology; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100449

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 localization in response to certain stimuli. Western blot analysis allows the quantification of changes in the distribution of these proteins between the cytoplasm and nucleus, whereas the posttranslational modifications specific to each compartment 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). [Display omitted] •A simple way to obtain tagged proteins in Xenopus embryos•Rapid separation of Xenopus embryonic cells into nuclear, cytoplasmic fractions•Quantifying the distribution of shuttle proteins between the cytoplasm and nucleus•Protein posttranslational modifications specific to the cytoplasm and nucleus can be studied 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 localization in response to certain stimuli. Western blot analysis allows the quantification of changes in the distribution of these proteins between the cytoplasm and nucleus, while the posttranslational modifications specific to each compartment can be identified by changes in electrophoretic mobility.