Preparation of cross-linked cellular extracts with formaldehyde

© 2014 Cold Spring Harbor Laboratory Press.

Bibliographic Details
Published in:Cold Spring Harbor protocols, Vol. 2014, No. 9 (2014), p. 1001-3
Main Author: Nilsen, Timothy W
Format: electronic Article
Language:English
ISSN:1559-6095
Item Description:Date Completed 23.06.2015
Date Revised 03.09.2014
published: Electronic
Citation Status MEDLINE
Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Physical Description:Online-Ressource
DOI:10.1101/pdb.prot080879
Subjects:
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Description:
  • © 2014 Cold Spring Harbor Laboratory Press.
  • Formaldehyde is a powerful cross-linking agent that elicits protein-protein and protein-nucleic acid cross-links. This protocol describes the formaldehyde cross-linking of intact cells followed by either the preparation of a whole-cell extract by sonication or the preparation of nuclear and cytoplasmic extracts by fractionation. Cross-linked extracts are treated with high salt or sodium dodecyl sulfate (SDS) to disrupt non-cross-linked aggregates. Protein-protein and protein-nucleic acid interactions can then be studied by purifying the components of interest from the extracts (e.g., using immunoprecipitation). Formaldehyde cross-links are reversible by heat