Micro Chromatin Immunoprecipitation (μChIP) from Early Mammalian Embryos
Micro Chromatin Immunoprecipitation (μChIP) from Early Mammalian Embryos
复制标题
DOI：
10.1007/978-1-4939-1594-1_17
复制
发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Klungland, Arne
中科院分区：
文献类型：
--
作者：
Dahl, John Arne;Klungland, Arne
染色质免疫沉淀（ChIP）是一种用于绘制体内蛋白质-DNA相互作用的强有力的方法。组蛋白修饰、转录因子和其他调节蛋白的基因组定位可以通过ChIP来揭示。然而，传统的ChIP方案需要使用大量的细胞，这阻碍了ChIP应用于稀有细胞类型。我们已经开发了ChIP检测，适用于从小细胞数量中免疫沉淀组蛋白或转录因子。在这里，我们描述了一种快速，但敏感的微（亩）ChIP协议产生高信噪比输出，适合于少至100个细胞。本章提供了一个详细的协议，从早期哺乳动物胚胎，也适用于任何样品的细胞数量有限的μ ChIP。这种优化的高信噪比ChIP方案的微小修改使其成为用于任何细胞数量的可靠工具（100 - 10（7））
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 (mu) ChIP protocol producing high signal to noise ratio output, suitable for as few as 100 cells. This chapter provides a detailed protocol for mu ChIP 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))