Assaying Chromatin Sirtuins

Abstract

Most of the sirtuins’ nuclear substrates identifi ed so far are histones or other chromatin-associated proteins and, thus, it is of special relevance the development of good biochemical techniques to analyze the biology of these proteins in the context of chromatin. Here, we describe several of the chromatin-based techniques to identify sirtuins’ substrates, including a chromatin immunoprecipitation (ChIP) protocol, an acid- extraction protocol, and a nucleosomal immunoprecipitation protocol to analyze putative sirtuin chromatin interactors.