Nuclear Import in Digitonin‐Permeabilized Cells
Open Access
- 1 January 2000
- journal article
- unit
- Published by Wiley in Current Protocols in Cell Biology
- Vol. 5 (1) , 11.7.1-11.7.17
- https://doi.org/10.1002/0471143030.cb1107s05
Abstract
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.Keywords
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