Protocol for label-free ribosome-associated protein enrichment from mammalian cells by RAPIDASH
Year: 2025
Type: article
Abstract: Ribosome-associated proteins (RAPs) enable modulation of gene expression at the level of mRNA translation in response to cellular needs. Here, we describe a method called ribosome-associated protein identification by affinity to sulfhydryl-charged resin (RAPIDASH) for tag-free isolation of RAP-bound ribosomes from mammalian samples for mass spectrometry-based proteomics. Samples are first lysed and then undergo sucrose cushion ultracentrifugation and subsequent chromatography using a sulfhydryl-charged resin. While RAPIDASH is optimized for mammalian samples, we expect that it can be adapted for non-mammalian samples. For complete details on the use and execution of this protocol, please refer to Susanto et al. 1
Source: STAR Protocols
Institutions: Stanford Medicine, Stanford University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
Funders: Damon Runyon Cancer Research Foundation, Stanford University, School of Medicine, Oregon Health and Science University, Agency for Science, Technology and Research, National Institutes of Health, School of Medicine, Stanford University, National Institute of Mental Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development
OpenAlex ID: W4413888022