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Optimized protocols for chromatin immunoprecipitation of exogenously expressed epitope-tagged proteins.
Fang, Wentong; Liao, Chengheng; Zhang, Qing.
Afiliação
  • Fang W; Department of Pharmacy, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
  • Liao C; Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: chengheng.liao@utsouthwestern.edu.
  • Zhang Q; Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: qing.zhang@utsouthwestern.edu.
STAR Protoc ; 4(1): 102050, 2023 03 17.
Article em En | MEDLINE | ID: mdl-36853721
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.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Fatores de Transcrição / Proteínas de Ligação a DNA Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Fatores de Transcrição / Proteínas de Ligação a DNA Idioma: En Ano de publicação: 2023 Tipo de documento: Article