Unit

UNIT 11.7 Nuclear Import in Digitonin-Permeabilized Cells

  1. Mary Shannon Moore,
  2. Eric D. Schwoebel

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb1107s05

Current Protocols in Cell Biology

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.

Author Information

  1. Baylor College of Medicine, Houston, Texas

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 2000

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.