title: RNA-binding Protein Immunoprecipitation (RIP) to Examine AUF1 Binding to Senescence-Associated Secretory Phenotype (SASP) Factor mRNA.
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 Alspachet al. (2014).
authors.affiliation: Departments of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, USA; ICCE Institute, Washington University School of Medicine, St. Louis, USA.
authors.forename: Elise
authors.initials: E
authors.lastname: Alspach
authors.affiliation: Departments of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, USA; Departments of Medicine, Washington University School of Medicine, St. Louis, USA; ICCE Institute, Washington University School of Medicine, St. Louis, USA.
authors.forename: Sheila A
authors.initials: SA
authors.lastname: Stewart