Protocol for chromatin immunoprecipitation of histone modifications in frozen adipose tissue

冷冻脂肪组织中组蛋白修饰的染色质免疫沉淀实验方案

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作者:Akin Cayir ,Tone M Tannæs ,Sadia Saeed ,Matthias Blüher ,Yvonne Böttcher

Abstract

Chromatin immunoprecipitation (ChIP) combined with sequencing has revolutionized our understanding of gene regulation; however, its application to frozen adipose tissue presents unique challenges due to the high levels of lipid content. Here, we present a protocol for ChIP of histone modifications in human frozen adipose tissue. We describe steps for tissue preparation, chromatin isolation, sonication, pre-clearing of chromatin, and immunoprecipitation. We then detail procedures for elution, crosslink reversal, chromatin purification, quality control, and library synthesis. Keywords: Chromatin immunoprecipitation (ChIP); Health Sciences.

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