Isolation of Peroxisomes from Yeast

Peroxisomes are multifunctional, dynamic organelles present in nearly all eukaryotic cells. Determining their structural and functional characteristics often requires obtaining isolated and purified peroxisomes via subcellular fractionation. Subcellular fractionation techniques are generally based on a three-step procedure: preparation of a cell-free homogenate (postnuclear supernatant), generation of an organellar pellet by differential centrifugation, and density gradient centrifugation. Here we introduce methods for small-scale isolation of peroxisomes from yeast cells using different gradient media as well as large-scale purification using a two-step gradient centrifugation.