A rapid micro chromatin immunoprecipitation assay (μChIP)
A rapid micro chromatin immunoprecipitation assay (μChIP)
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DOI：
10.1038/nprot.2008.68
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Collas, Philippe
中科院分区：
文献类型：
--
作者：
Dahl, John Arne;Collas, Philippe
蛋白质与DNA的相互作用介导了许多关键的核功能。染色质免疫沉淀（ChIP）是一种研究蛋白质- dna相互作用的强大技术。然而，目前的ChIP检测要么需要大量的细胞数量，这阻碍了它们在罕见细胞样本或小组织活检中的应用，要么涉及冗长的程序。我们在这里描述了一个为期1天的微芯片（mu ChIP）方案，适用于从单个批次的1000个细胞中进行多达8次平行组蛋白和/或转录因子免疫沉淀。mu ChIP技术也适用于在100个细胞中监测一种蛋白质与多个基因组位点的关联。交联和染色质制备步骤的改变也使mu ChIP适用于类似于1毫米(3)新鲜或冷冻组织活检。从细胞固定到pcr准备的DNA， 16个芯片的过程大约需要8小时。
Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or involve lengthy procedures. We describe here a 1-day micro ChIP (mu ChIP) protocol suitable for up to eight parallel histone and/or transcription factor immunoprecipitations from a single batch of 1,000 cells. mu ChIP technique is also suitable for monitoring the association of one protein with multiple genomic sites in 100 cells. Alterations in cross-linking and chromatin preparation steps also make mu ChIP applicable to similar to 1-mm(3) fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes similar to 8 h for 16 ChIPs.