染色质
组蛋白修饰酶
生物
组蛋白
化学
生物化学
组蛋白密码
组蛋白甲基转移酶
分子生物学
细胞生物学
DNA
核小体
作者
David Shechter,Holger L. Dormann,C. David Allis,Sandra B. Hake
出处
期刊:Nature Protocols
[Springer Nature]
日期:2007-06-01
卷期号:2 (6): 1445-1457
被引量:940
标识
DOI:10.1038/nprot.2007.202
摘要
Histone proteins are the major protein components of chromatin, the physiologically relevant form of the genome (or epigenome) in all eukaryotic cells. Chromatin is the substrate of many biological processes, such as gene regulation and transcription, replication, mitosis and apoptosis. Since histones are extensively post-translationally modified, the identification of these covalent marks on canonical and variant histones is crucial for the understanding of their biological significance. Many different biochemical techniques have been developed to purify and separate histone proteins. Here, we present standard protocols for acid extraction and salt extraction of histones from chromatin; separation of extracted histones by reversed-phase HPLC; analysis of histones and their specific post-translational modification profiles by acid urea (AU) gel electrophoresis and the additional separation of non-canonical histone variants by triton AU(TAU) and 2D TAU electrophoresis; and immunoblotting of isolated histone proteins with modification-specific antibodies.
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