Dahl John Arne, Collas Philippe
Institute of Basic Medical Sciences, Department of Biochemistry, Faculty of Medicine, University of Oslo, Blindern, Oslo, Norway.
Front Biosci. 2007 Sep 1;12:4925-31. doi: 10.2741/2438.
Chromatin immunoprecipitation (ChIP) is to date a technique of choice for studying protein-DNA interactions. ChIP has been used for mapping the location of modified histones on DNA, often in relation to transcription or differentiation. A drawback of current ChIP protocols, however, is the requirement for large cell numbers, which limits the applicability of ChIP to rare cell samples. The procedure is also tedious and time consuming. We systematically evaluated and modified critical steps in the ChIP procedure to develop a quick and quantitative ChIP assay, referred to as Q2ChIP, suitable for up to 1,000 histone immunoprecipitations or 100 transcription factor immunoprecipitations from as few as 100,000 cells under cross-linking conditions. Analysis of the precipitated DNA by real time PCR enables quantification of the relative amount of a specific histone modification or transcription factor associated with a specific locus. This communication describes all steps of the Q2ChIP procedure as it is carried out in our laboratory.
染色质免疫沉淀法（ChIP）是目前研究蛋白质 - DNA相互作用的首选技术。ChIP已被用于绘制DNA上修饰组蛋白的位置，这通常与转录或分化有关。然而，当前ChIP方案的一个缺点是需要大量细胞，这限制了ChIP在稀有细胞样本中的适用性。该过程也很繁琐且耗时。我们系统地评估并修改了ChIP过程中的关键步骤，以开发一种快速定量的ChIP检测方法，称为Q2ChIP，适用于在交联条件下从低至100,000个细胞中进行多达1000次组蛋白免疫沉淀或100次转录因子免疫沉淀。通过实时PCR对沉淀的DNA进行分析，可以定量与特定基因座相关的特定组蛋白修饰或转录因子的相对量。本通讯描述了我们实验室中进行的Q2ChIP过程的所有步骤。
