A streamlined method for rapid and sensitive chromatin immunoprecipitation

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• 作者：Michael L. Sikes ; Justin M. Bradshaw ; Wendell T. Ivory ; Jessica L. Lunsford ; Ruth E. McMillan ; Clayton R. Morrison
• 关键词：Chromatin immunoprecipitation ; Histone ; Epigenetics ; cad ; TEA ; Thymocyte ; Rag
• 刊名：Journal of Immunological Methods
• 出版年：2009
• 出版时间：15 May 2009
• 年：2009
• 卷：344
• 期：1
• 页码：58-63
• 全文大小：286 K

文摘

We report a streamlined procedure to efficiently carry samples from chromatin to qPCR-compatible DNA in as little as 4 h. We use this streamlined ChIP to quantify histone H3 modifications at active (cad) and repressed (T early alpha) promoters in a Rag1-deficient pro-T cell line after 1–2 h IP. We further show that the protocol readily quantified histone modifications in chromatin from 10⁴ Rag-deficient DN thymocytes. Taken together, these data outline a simple, cost-effective procedure for efficient ChIP analysis.