A non-isotopic in vitro assay for histone acetylation

David Kuninger, James Lundblad, Anthony Semirale, Peter Rotwein

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

We describe a simple, robust, and relatively inexpensive non-radioactive in vitro assay for measuring histone acetyl-transferase activity. The assay takes advantage of easy to purify recombinant E. coli-derived fusion proteins containing the NH2-terminal tails of histones H3 and H4 linked to epitope-tagged maltose-binding protein (MBP), and immunoblotting with antibodies specific to acetylated H3 and H4. Here we show the specificity and dynamic range of this assay for the histone acetyl-transferases, p300 and PCAF. This assay may be adapted readily for other substrates by simply generating new fusion proteins and for other acetyl-transferases by modifying reaction conditions.

Original languageEnglish (US)
Pages (from-to)253-260
Number of pages8
JournalJournal of Biotechnology
Volume131
Issue number3
DOIs
StatePublished - Sep 15 2007

Keywords

  • Acetyl-transferase assay
  • Histone H3
  • Histone H4
  • Non-isotopic
  • PCAF
  • p300

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'A non-isotopic in vitro assay for histone acetylation'. Together they form a unique fingerprint.

  • Cite this