Dahl John Arne, Klungland Arne
Institute of Clinical Medicine, Division of diagnostics and intervention, Department of Microbiology, Oslo university hospital, Rikshospitalet, NO-0027, Oslo, Norway,
Methods Mol Biol. 2015;1222:227-45. doi: 10.1007/978-1-4939-1594-1_17.
Chromatin immunoprecipitation (ChIP) is a powerful method for mapping protein-DNA interactions in vivo. Genomic localization of histone modifications, transcription factors, and other regulatory proteins can be revealed by ChIP. However, conventional ChIP protocols require the use of large numbers of cells, which prevents the application of ChIP to rare cell types. We have developed ChIP assays suited for the immunoprecipitation of histone proteins or transcription factors from small cell numbers. Here we describe a rapid, yet sensitive micro (μ)ChIP protocol producing high signal to noise ratio output, suitable for as few as 100 cells. This chapter provides a detailed protocol for μChIP from early mammalian embryos, also suitable for any sample of limited numbers of cells. Minor modifications of this optimized high signal to noise ChIP protocol make it a reliable tool for the use with any cell number (100-10(7)).
染色质免疫沉淀（ChIP）是一种在体内绘制蛋白质 - DNA 相互作用图谱的强大方法。通过 ChIP 可以揭示组蛋白修饰、转录因子和其他调控蛋白的基因组定位。然而，传统的 ChIP 方案需要使用大量细胞，这使得 ChIP 无法应用于稀有细胞类型。我们开发了适用于从小细胞数量中免疫沉淀组蛋白或转录因子的 ChIP 分析方法。在此，我们描述一种快速且灵敏的微量（μ）ChIP 方案，该方案能产生高信噪比输出，适用于少至 100 个细胞。本章提供了从早期哺乳动物胚胎进行 μChIP 的详细方案，也适用于任何细胞数量有限的样本。对这种优化的高信噪比 ChIP 方案进行微小修改，使其成为适用于任何细胞数量（100 - 10⁷）的可靠工具。
