甲醛交联嗅感觉神经元的染色质免疫沉淀。

Chromatin Immunoprecipitation from Formaldehyde Cross-Linked Olfactory Sensory Neurons.

机构信息

Department of Molecular Biology and Biochemistry, Nelson Biological Laboratories, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.

出版信息

Methods Mol Biol. 2023;2710:71-82. doi: 10.1007/978-1-0716-3425-7_6.

DOI:10.1007/978-1-0716-3425-7_6

PMID:37688725

Abstract

Chromatin immunoprecipitation (ChIP) allows a researcher to determine the genomic occupancy of nuclear proteins, providing insight into the roles of transcription factors, chromatin modifiers, histone modifications, and other factors bound to DNA. Protein-DNA interactions are first fixed in vivo by chemical cross-linking, and then a target protein is captured together with any associated DNA by an antibody mediated pull-down. The co-immunoprecipitated DNA can then be assayed by quantitative PCR or deep sequencing. Here, we demonstrate this technique using murine olfactory sensory neurons (OSNs) purified using fluorescence-activated cell sorting (FACS) and antibodies for the ubiquitous chromatin protein CTCF.

摘要

染色质免疫沉淀（ChIP）可让研究人员确定核蛋白在基因组上的位置，从而深入了解转录因子、染色质修饰因子、组蛋白修饰以及与 DNA 结合的其他因子的作用。首先，通过化学交联将蛋白质 - DNA 相互作用在体内固定，然后通过抗体介导的下拉法将靶蛋白与其任何相关 DNA 一起捕获。随后，可以通过定量 PCR 或深度测序来检测共免疫沉淀的 DNA。在这里，我们使用通过荧光激活细胞分选（FACS）纯化的小鼠嗅觉感觉神经元（OSN）和用于普遍存在的染色质蛋白 CTCF 的抗体演示了该技术。