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Author:
Date:
2015-05-20
[Abstract] Immunoprecipitation and subsequent isolation of nucleic acids allows for the investigation of protein:nucleic acid interactions. RNA-binding protein immunoprecipitation (RIP) is used for the analysis of protein interactions with mRNA. Combining RIP with quantitative real-time PCR (qRT-PCR) further enhances the RIP technique by allowing for the quantitative assessment of RNA-binding protein interactions with their target mRNAs, and how these interactions change in different cellular settings. Here, we describe the immunoprecipitation of the RNA-binding protein AUF1 with several different factors associated with the senescence-associated secretory phenotype (SASP) (Alspach and Stewart, 2013), specifically IL6 and IL8. This protocol was originally published in Alspach et al. (2014).
[摘要] 免疫沉淀和随后的核酸分离允许研究蛋白质：核酸相互作用。 RNA结合蛋白免疫沉淀（RIP）用于分析蛋白质与mRNA的相互作用。 结合RIP与定量实时PCR（qRT-PCR）通过允许RNA结合蛋白与其靶mRNA的相互作用的定量评估以及这些相互作用在不同细胞设置中如何变化来进一步增强RIP技术。 在这里，我们描述RNA结合蛋白AUF1与几种不同的因素与衰老相关的分泌表型（SASP）（Alspach和斯图尔特，2013年），特别是IL6和IL8相关的免疫沉淀。 该协议最初发表在Alspach等人（2014）。
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