Optimized protocols for chromatin immunoprecipitation of exogenously expressed epitope-tagged proteins

针对外源表达的表位标记蛋白的染色质免疫沉淀,优化了相关实验方案。

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作者:Wentong Fang ,Chengheng Liao ,Qing Zhang

Abstract

Chromatin immunoprecipitation (ChIP) assay is widely used for investigating the interaction between DNA and DNA-binding proteins such as transcription factors, co-factors, or chromatin-associated proteins. However, a successful ChIP assay largely depends on the quality of a ChIP-grade primary antibody. In cases where specific antibodies are unavailable or with low binding affinity, here, we describe a tailored protocol to achieve robust and reproducible chromatin binding by expressing an exogenous epitope-tagged protein in cells, followed by ChIP assays using a tag-specific antibody. For complete details on the use and execution of this protocol, please refer to Fang et al. (2021)1 and Kidder et al. (2011).2. Keywords: Cell Biology; Genomics; Molecular Biology; RNAseq.

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