Isolation of Autophagy Competent Lysosomes from Cancer, Cells by Differential Large-Scale, Multilayered Density Gradient Centrifugations

Isolation of Autophagy Competent Lysosomes from Cancer


Accurate isolation of functional and intact lysosomes enables the quantification and analyses of abundances, dynamic changes and enrichment levels of lysosomal content, allowing specific lysosomal investigations induced by autophagy. In this protocol chapter, we describe detailed practical instructions and advices for an efficacious lysosomal enrichment and isolation procedure by differential multilayered density gradient centrifugations using human cancer cell lines. By this method, intact and autophagy competent lysosomes can be isolated from cancer cells based on their distinct density and obtained fractions can further be analyzed for functional lysosomal assays, as well as for protein or metabolic loads to identify select spatiotemporal changes by comparative quantitative measurement. This method has been used to enrich lysosomes from a variety of cancer cells with activated chaperone-mediated autophagy, but can be optimized for other cell lines and tissues for multiple autophagy-induced conditions.

Вас заинтересует / Intresting for you:

A Balance Between Autophagy an...
A Balance Between Autophagy an... 917 views Дэйзи ак-Макарова Mon, 21 Feb 2022, 07:31:06
NON-Fungible Token (NFT) Histo...
NON-Fungible Token (NFT) Histo... 396 views Zero Cool Wed, 09 Feb 2022, 18:20:07
NON-FUNGIBLE TOKEN (NFT) mean... 871 views Zero Cool Wed, 09 Feb 2022, 18:22:00
How to create NFTs (NON-Fungib...
How to create NFTs (NON-Fungib... 518 views Zero Cool Thu, 10 Feb 2022, 13:53:20
Comments (0)
There are no comments posted here yet
Leave your comments
Posting as Guest
Suggested Locations