An easy assay for histone acetyltransferase activity using a PhosphorImager.
A simple radiometric assay for histone acetyltransferase (HAT) activity employing a PhosphorImager is described. In the proposed procedure, following incubation of [1-(14)C]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.
Poveda
Ana
A
Departament de Bioquímica i Biologia Molecular, Facultat de Ciències Biològiques, 46100 Burjassot (València), Spain.
Sendra
Ramon
R
D000818:Animals; D051548:Histone Acetyltransferases / Q000032:analysis* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D007202:Indicators and Reagents / Q000737:chemistry; D011874:Radiometry / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D006657:Histones; D007202:Indicators and Reagents; D051548:Histone Acetyltransferases