UNIT 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Abmayr, S. M. and Workman, J. L. 2001. Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells. Current Protocols in Molecular Biology. 12:12.1.
- Published Online: 1 MAY 2001
- Published Print: OCT 1993
This is not the most recent version of the article. View current version (1 AUG 2006)
Extracts prepared from the isolated nuclei of cultured cells are functional in accurate in vitro transcription and mRNA processing. Thus, such extracts can be used directly for functional studies and as the starting material for purification of the proteins involved in these processes. In this unit, reproducible extracts from different preparations of nuclei are generated. The conditions described are optimized to produce transcriptionally active extracts from HeLa cells. A describes how to optimize the basic procedure to increase the yield of specific proteins or activities from different cell types, and a second describes the preparation of the cytoplasmic (S-100) fraction.