果蝇胚胎的染色质制备及染色质免疫沉淀

Chromatin Preparation and Chromatin Immuno-precipitation from Drosophila Embryos.

作者信息

Löser Eva, Latreille Daniel, Iovino Nicola

机构信息

Max Planck Institute of Immunobiology and Epigenetics, Stübeweg 51, Freiburg, 79108, Germany.

出版信息

Methods Mol Biol. 2016;1480:23-36. doi: 10.1007/978-1-4939-6380-5_3.

DOI:10.1007/978-1-4939-6380-5_3

PMID:27659972

Abstract

This protocol provides specific details on how to perform Chromatin immunoprecipitation (ChIP) from Drosophila embryos. ChIP allows the matching of proteins or histone modifications to specific genomic regions. Formaldehyde-cross-linked chromatin is isolated and antibodies against the target of interest are used to determine whether the target is associated with a specific DNA sequence. This can be performed in spatial and temporal manner and it can provide information about the genome-wide localization of a given protein or histone modification if coupled with deep sequencing technology (ChIP-Seq).

摘要

本方案提供了关于如何从果蝇胚胎中进行染色质免疫沉淀（ChIP）的具体细节。ChIP可将蛋白质或组蛋白修饰与特定基因组区域进行匹配。分离甲醛交联的染色质，并使用针对目标蛋白的抗体来确定该目标蛋白是否与特定DNA序列相关联。这可以在空间和时间维度上进行，如果与深度测序技术（ChIP-Seq）相结合，它可以提供有关给定蛋白质或组蛋白修饰在全基因组范围内定位的信息。