Unit

UNIT 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells

  1. Susan M. Abmayr,
  2. Jerry L. Workman

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1201s24

Current Protocols in Molecular Biology

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.

Author Information

  1. Pennsylvania State University, University Park, Pennsylvania

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1993

This is not the most recent version of the article. View current version (1 AUG 2006)

Abstract

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 support protocol describes how to optimize the basic procedure to increase the yield of specific proteins or activities from different cell types, and a second support protocol describes the preparation of the cytoplasmic (S-100) fraction.