UNIT 11.7 Nuclear Import in Digitonin-Permeabilized Cells
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Moore, M. S. and Schwoebel, E. D. 2001. Nuclear Import in Digitonin-Permeabilized Cells. Current Protocols in Cell Biology. 5:11.7:11.7.1–11.7.17.
- Published Online: 1 MAY 2001
- Published Print: JAN 2000
Import of proteins into the nucleus of a cell is a complex process that can be reconstituted in vitro. Digitonin-permeabilized cells are washed free of cytosolic factors to provide competent nuclei. Cytosolic factors for import are provided by an extract of Xenopus ovarian cells. Fluorochrome-conjugated probes, cloned proteins fused to green fluorescent protein, or antibodies to the protein of interest are used to visualize nuclear import. The system can also be used to identify nuclear localization sequences (NLS) of imported proteins.