Proteomics

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A PEROXO-Tag enables rapid isolation of peroxisomes from human cells

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 one hour 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 minutes 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.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: Elizabeth Boydston  

LAB HEAD: Sebastian Lourido

PROVIDER: PXD018918 | Pride | 2020-05-20

REPOSITORIES: Pride

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20191124_GJR_exp171_IP_Pool.mzML Mzml
20191124_GJR_exp171_IP_Pool.pride.mgf.gz Mgf
GJRexp171_WholeCell_Pool-(1).pride.mgf.gz Mgf
GJRexp171_WholeCell_Pool-1.mzML Mzml
Human_Uniprot_Sprt_Trembl_Isoforms_Release2019_01_Contams_20190205.fasta Fasta
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A PEROXO-Tag Enables Rapid Isolation of Peroxisomes from Human Cells.

Ray G Jordan GJ   Boydston Elizabeth A EA   Shortt Emily E   Wyant Gregory A GA   Lourido Sebastian S   Chen Walter W WW   Sabatini David M DM  

iScience 20200428 5

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  ...[more]

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