A PEROXO-Tag Enables Rapid Isolation of Peroxisomes from Human Cells.
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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. |
Year of Publication | 2020
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Journal | iScience
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Volume | 23
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Issue | 5
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Pages | 101109
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Date Published | 2020 May 22
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ISSN | 2589-0042
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DOI | 10.1016/j.isci.2020.101109
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PubMed ID | 32417403
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