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A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting

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Standard

A Proteomics Approach to the Protein Normalization Problem : Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting. / Wiśniewski, Jacek R; Mann, Matthias.

I: Journal of Proteome Research, Bind 15, Nr. 7, 01.07.2016, s. 2321-6.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Wiśniewski, JR & Mann, M 2016, 'A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting', Journal of Proteome Research, bind 15, nr. 7, s. 2321-6. https://doi.org/10.1021/acs.jproteome.6b00403

APA

Wiśniewski, J. R., & Mann, M. (2016). A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting. Journal of Proteome Research, 15(7), 2321-6. https://doi.org/10.1021/acs.jproteome.6b00403

Vancouver

Wiśniewski JR, Mann M. A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting. Journal of Proteome Research. 2016 jul 1;15(7):2321-6. https://doi.org/10.1021/acs.jproteome.6b00403

Author

Wiśniewski, Jacek R ; Mann, Matthias. / A Proteomics Approach to the Protein Normalization Problem : Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting. I: Journal of Proteome Research. 2016 ; Bind 15, Nr. 7. s. 2321-6.

Bibtex

@article{e0f0dd353a8d4930954f5bc544017f5e,
title = "A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting",
abstract = "Proteomics and other protein-based analysis methods such as Western blotting all face the challenge of discriminating changes in the levels of proteins of interest from inadvertent changes in the amount loaded for analysis. Mass-spectrometry-based proteomics can now estimate the relative and absolute amounts of thousands of proteins across diverse biological systems. We reasoned that this new technology could prove useful for selection of very stably expressed proteins that could serve as better loading controls than those traditionally employed. Large-scale proteomic analyses of SDS lysates of cultured cells and tissues revealed deglycase DJ-1 as the protein with the lowest variability in abundance among different cell types in human, mouse, and amphibian cells. The protein constitutes 0.069 ± 0.017{\%} of total cellular protein and occurs at a specific concentration of 34.6 ± 8.7 pmol/mg of total protein. Since DJ-1 is ubiquitous and therefore easily detectable with several peptides, it can be helpful in normalization of proteomic data sets. In addition, DJ-1 appears to be an advantageous loading control for Western blot that is superior to those used commonly used, allowing comparisons between tissues and cells originating from evolutionarily distant vertebrate species. Notably, this is not possible by the detection and quantitation of housekeeping proteins, which are often used in the Western blot technique. The approach introduced here can be applied to select the most appropriate loading controls for MS-based proteomics or Western blotting in any biological system.",
keywords = "Animals, Blotting, Western, Genes, Essential, Humans, Mass Spectrometry, Protein Deglycase DJ-1, Proteins, Proteomics, Sample Size, Journal Article",
author = "Wiśniewski, {Jacek R} and Matthias Mann",
year = "2016",
month = "7",
day = "1",
doi = "10.1021/acs.jproteome.6b00403",
language = "English",
volume = "15",
pages = "2321--6",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "7",

}

RIS

TY - JOUR

T1 - A Proteomics Approach to the Protein Normalization Problem

T2 - Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting

AU - Wiśniewski, Jacek R

AU - Mann, Matthias

PY - 2016/7/1

Y1 - 2016/7/1

N2 - Proteomics and other protein-based analysis methods such as Western blotting all face the challenge of discriminating changes in the levels of proteins of interest from inadvertent changes in the amount loaded for analysis. Mass-spectrometry-based proteomics can now estimate the relative and absolute amounts of thousands of proteins across diverse biological systems. We reasoned that this new technology could prove useful for selection of very stably expressed proteins that could serve as better loading controls than those traditionally employed. Large-scale proteomic analyses of SDS lysates of cultured cells and tissues revealed deglycase DJ-1 as the protein with the lowest variability in abundance among different cell types in human, mouse, and amphibian cells. The protein constitutes 0.069 ± 0.017% of total cellular protein and occurs at a specific concentration of 34.6 ± 8.7 pmol/mg of total protein. Since DJ-1 is ubiquitous and therefore easily detectable with several peptides, it can be helpful in normalization of proteomic data sets. In addition, DJ-1 appears to be an advantageous loading control for Western blot that is superior to those used commonly used, allowing comparisons between tissues and cells originating from evolutionarily distant vertebrate species. Notably, this is not possible by the detection and quantitation of housekeeping proteins, which are often used in the Western blot technique. The approach introduced here can be applied to select the most appropriate loading controls for MS-based proteomics or Western blotting in any biological system.

AB - Proteomics and other protein-based analysis methods such as Western blotting all face the challenge of discriminating changes in the levels of proteins of interest from inadvertent changes in the amount loaded for analysis. Mass-spectrometry-based proteomics can now estimate the relative and absolute amounts of thousands of proteins across diverse biological systems. We reasoned that this new technology could prove useful for selection of very stably expressed proteins that could serve as better loading controls than those traditionally employed. Large-scale proteomic analyses of SDS lysates of cultured cells and tissues revealed deglycase DJ-1 as the protein with the lowest variability in abundance among different cell types in human, mouse, and amphibian cells. The protein constitutes 0.069 ± 0.017% of total cellular protein and occurs at a specific concentration of 34.6 ± 8.7 pmol/mg of total protein. Since DJ-1 is ubiquitous and therefore easily detectable with several peptides, it can be helpful in normalization of proteomic data sets. In addition, DJ-1 appears to be an advantageous loading control for Western blot that is superior to those used commonly used, allowing comparisons between tissues and cells originating from evolutionarily distant vertebrate species. Notably, this is not possible by the detection and quantitation of housekeeping proteins, which are often used in the Western blot technique. The approach introduced here can be applied to select the most appropriate loading controls for MS-based proteomics or Western blotting in any biological system.

KW - Animals

KW - Blotting, Western

KW - Genes, Essential

KW - Humans

KW - Mass Spectrometry

KW - Protein Deglycase DJ-1

KW - Proteins

KW - Proteomics

KW - Sample Size

KW - Journal Article

U2 - 10.1021/acs.jproteome.6b00403

DO - 10.1021/acs.jproteome.6b00403

M3 - Journal article

C2 - 27297043

VL - 15

SP - 2321

EP - 2326

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 7

ER -

ID: 186876564