UNIT 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells

  1. Susan M. Abmayr,
  2. Tingting Yao,
  3. Tari Parmely,
  4. Jerry L. Workman

Published Online: 1 AUG 2006

DOI: 10.1002/0471142727.mb1201s75

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Abmayr, S. M., Yao, T., Parmely, T. and Workman, J. L. 2006. Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells. Current Protocols in Molecular Biology. 75:12.1:12.1.1–12.1.10.

Author Information

  1. The Stowers Institute for Medical Research, Kansas City, Missouri

Publication History

  1. Published Online: 1 AUG 2006
  2. Published Print: JUL 2006


Extracts prepared from the isolated nuclei of cultured cells have been instrumental in dissecting the mechanisms by which transcription and mRNA processing occur. These extracts are able to recapitulate accurate transcription initiation and splicing in vitro, which has been useful in direct functional studies. They also serve as the starting material for purification of proteins that can then be reassembled in functional studies or examined in more detail biochemically. This unit describes the preparation of nuclear extracts from cultured cells and optimized production of transcriptionally active extracts from HeLa cells. Additional protocols describe optimization of the method to increase the yield of specific proteins, adaptation of the method for downstream applications such as affinity purification, and preparation of the cytoplasmic (S-100) fraction.


  • Nuclear extract;
  • cytoplasmic extract