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小鼠海马体细胞和组织中的染色质免疫沉淀

Chromatin immunoprecipitation in mouse hippocampal cells and tissues.

作者信息

Sailaja Badi Sri, Takizawa Takumi, Meshorer Eran

机构信息

Department of Genetics, Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.

出版信息

Methods Mol Biol. 2012;809:353-64. doi: 10.1007/978-1-61779-376-9_24.

Abstract

Chromatin immunoprecipitation (ChIP) has been developed for studying protein-DNA interactions and has been extensively used for mapping the localization of posttranslationally modified histones, histone variants, transcription factors, or chromatin modifying enzymes at a given locus or on a genome-wide scale. ChIP methods have been modified and improved over the years to fit a variety of different cell types and tissues. Here, we present a detailed protocol for hippocampal ChIP, of both minced tissue and enzyme-separated hippocampal cells. This protocol enables to study chromatin-protein interactions in a specified population of hippocampal cells, allowing to study chromatin regulation in the central nervous system in a variety of conditions and disorders. Our assay has been developed for histone modifications but is suited for any chromatin binding protein for which specific ChIP-grade antibodies are available.

摘要

染色质免疫沉淀(ChIP)技术已被开发用于研究蛋白质与DNA的相互作用,并已广泛应用于在给定基因座或全基因组范围内绘制翻译后修饰的组蛋白、组蛋白变体、转录因子或染色质修饰酶的定位图谱。多年来,ChIP方法不断得到改进和完善,以适应各种不同的细胞类型和组织。在这里,我们提供了一个详细的海马ChIP实验方案,包括切碎的组织和酶分离的海马细胞。该方案能够研究特定海马细胞群体中的染色质-蛋白质相互作用,从而可以在各种条件和疾病下研究中枢神经系统中的染色质调控。我们的检测方法是针对组蛋白修饰开发的,但适用于任何有特异性ChIP级抗体的染色质结合蛋白。

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