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A PEROXO-Tag Enables Rapid Isolation of Peroxisomes from Human Cells.

Authors :
Ray GJ
Boydston EA
Shortt E
Wyant GA
Lourido S
Chen WW
Sabatini DM
Source :
IScience [iScience] 2020 May 22; Vol. 23 (5), pp. 101109. Date of Electronic Publication: 2020 Apr 28.
Publication Year :
2020

Abstract

Peroxisomes are metabolic organelles that perform a diverse array of critical functions in human physiology. Traditional isolation methods for peroxisomes can take more than 1 h to complete and can be laborious to implement. To address this, we have now extended our prior work on rapid organellar isolation to peroxisomes via the development of a peroxisomally localized 3XHA epitope tag ("PEROXO-Tag") and associated immunoprecipitation ("PEROXO-IP") workflow. Our PEROXO-IP workflow has excellent reproducibility, is easy to implement, and achieves highly rapid (~10 min post homogenization) and specific isolation of human peroxisomes, which we characterize here via proteomic profiling. By offering speed, specificity, reproducibility, and ease of use, the PEROXO-IP workflow should facilitate studies on the biology of peroxisomes.<br />Competing Interests: Declaration of Interests The authors declare no competing interests.<br /> (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
2589-0042
Volume :
23
Issue :
5
Database :
MEDLINE
Journal :
IScience
Publication Type :
Academic Journal
Accession number :
32417403
Full Text :
https://doi.org/10.1016/j.isci.2020.101109