| Literature DB >> 33654706 |
Abel Cuevas-Bermúdez1, Ana I Garrido-Godino1, Francisco Gutiérrez-Santiago1, Francisco Navarro1,2.
Abstract
We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using zymolyase to obtain the spheroplast, which is employed in many other procedures. Employing small amount of culture cells and small volumes of solutions during the yChEFs procedure is very useful to allow many samples to be handled at the same time, and also reduces costs and efforts. The purified proteins associated with chromatin fractions obtained by yChEFs can be analyzed by Western blot (Figure 1) or combined with mass spectrometry for proteomic analyses.Entities:
Keywords: Chromatin; Mass spectrometry; Proteins; Saccharomyces cerevisiae; Yeast
Year: 2020 PMID: 33654706 PMCID: PMC7853979 DOI: 10.21769/BioProtoc.3471
Source DB: PubMed Journal: Bio Protoc ISSN: 2331-8325