Journal
ANALYTICAL BIOCHEMISTRY
Volume 383, Issue 2, Pages 296-300Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2008.08.029
Keywords
Acetylation; Acetyltransferase; Chromatin; Histones; PhosphorImager
Funding
- Ministerio de Educacion y Ciencia, Spain [BFU2005-02603]
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A simple radiometric assay for histone acetyltransferase (HAT) activity employing a PhosphorImager is described. In the proposed procedure, following incubation of [1-C-14]acetyl coenzyme A (CoA), histones, and HAT enzyme, radiolabeled histones are fixed on GF/F glass microfiber filter while the excess of acetyl CoA is washed out. Afterward, the filter is exposed to a phosphor-screen and the resulting spot signals are quantified with a PhosphorImager. Given the small volumes required, the new assay reduces reagent consumption and contaminated waste. Moreover, the assay can be performed with a large number of samples simultaneously, is applicable on different protein substrates, and is adaptable to the analysis of other protein modifications. (C) 2008 Elsevier Inc. All rights reserved.
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