Purification of Yeast Peroxisomes

Peroxisomes are ubiquitous subcellular organelles of eukaryotic cells. As with all organelles, peroxisomes can be purified from cell lysates using a combination of differential centrifugation and density gradient centrifugation. Here, we describe a method for purifying peroxisomes from the yeast Saccharomyces cerevisiae . The method involves gentle lysis of yeast spheroplasts, followed by differential centrifugation to obtain a crude organelle pellet enriched for peroxisomes and mitochondria. To separate peroxisomes from mitochondria, the organelle pellet is resuspended and spun through a sucrose density gradient. Peroxisomes purified in this way can be used to explore whether a protein of interest might be associated with the organelle. These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.