Chapter title |
Proteostasis
|
---|---|
Chapter number | 19 |
Book title |
Proteostasis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3756-1_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3754-7, 978-1-4939-3756-1
|
Authors |
Aguado, Carmen, Pérez-Jiménez, Eva, Lahuerta, Marcos, Knecht, Erwin, Carmen Aguado, Eva Pérez-Jiménez, Marcos Lahuerta, Erwin Knecht |
Editors |
Rune Matthiesen |
Abstract |
Lysosomes participate within the cells in the degradation of organelles, macromolecules, and a wide variety of substrates. In any study on specific roles of lysosomes, both under physiological and pathological conditions, it is advisable to include methods that allow their reproducible and reliable isolation. However, purification of lysosomes is a difficult task, particularly in the case of cultured cells. This is mainly because of the heterogeneity of these organelles, along with their low number and high fragility. Also, isolation methods, while disrupting plasma membranes, have to preserve the integrity of lysosomes, as the breakdown of their membranes releases enzymes that could damage all cell organelles, including themselves. The protocols described below have been routinely used in our laboratory for the specific isolation of lysosomes from rat liver, NIH/3T3, and other cultured cells, but can be adapted to other mammalian tissues or cell lines. |
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