Determination of CHK1 Cellular Localization by Immunofluorescence Microscopy

Yu-Che Cheng; Sheau-Yann Shieh

doi:10.1007/978-1-0716-1217-0_1

Determination of CHK1 Cellular Localization by Immunofluorescence Microscopy

Methods Mol Biol. 2021:2267:1-6. doi: 10.1007/978-1-0716-1217-0_1.

Authors

Yu-Che Cheng¹, Sheau-Yann Shieh²

Affiliations

¹ Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
² Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. sy88@ibms.sinica.edu.tw.

PMID: 33786781
DOI: 10.1007/978-1-0716-1217-0_1

Abstract

Many proteins involved in the DNA damage pathway shuttle between the cytoplasm and nucleus, and their localizations are important for functions. In that regard, immunofluorescence microscopy has been widely used to delineate the temporal and spatial regulation of proteins. Here, we describe an unconventional method for studying the cellular localization of CHK1, a cell cycle checkpoint kinase that undergoes shuttling from the cytoplasm to the nucleus in response to genotoxic stress. In this study, we included an acid extraction step to better reveal the nuclear localization of CHK1.

Keywords: CHK1; DNA damage response; HCl extraction; Immunofluorescence microscopy; Nuclear-cytoplasmic shuttling.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Checkpoint Kinase 1 / metabolism*
Fluorescent Antibody Technique / methods*
HCT116 Cells
Humans
Microscopy, Confocal / methods
Protein Transport

Substances

CHEK1 protein, human
Checkpoint Kinase 1