Isolation and Characterization of Lipid Droplets from Yeast

Lipid droplets are cell compartments serving as a depot for non-polar lipids, mainly triacylglycerols and steryl esters. Furthermore, these compartments are involved in lipid metabolism, signal transduction, membrane biosynthesis and trafficking, as well as storage of proteins, and additional functions may be discovered in the future. A crucial prerequisite for detailed studies of lipid droplets is the availability of a highly purified cell fraction. Here, a protocol for the isolation of lipid droplets from the model organism yeast Saccharomyces cerevisiae is presented. Characteristics of a lipid droplet fraction are a limited number of proteins and a high content of non-polar lipids. Most common problems occurring during isolation of lipid droplets and advices to overcome these problems are included.