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Purification of Yeast Peroxisomes

 Last Update: 2021-02-24
 Source: Internet
 Author: User

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density gradient centrifugation

density gradient

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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.

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