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Selected reaction monitoring for quantitative proteomics: a tutorial
Systems biology relies on data sets in which the same group of proteins is consistently identified and precisely quantified across multiple samples, a requirement that is only partially achieved by current proteomics approaches. Selected reaction monitoring (SRM)—also called multiple reaction monito...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
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Nature Publishing Group
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2583086/ https://www.ncbi.nlm.nih.gov/pubmed/18854821 http://dx.doi.org/10.1038/msb.2008.61 |
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author | Lange, Vinzenz Picotti, Paola Domon, Bruno Aebersold, Ruedi |
author_facet | Lange, Vinzenz Picotti, Paola Domon, Bruno Aebersold, Ruedi |
author_sort | Lange, Vinzenz |
collection | PubMed |
description | Systems biology relies on data sets in which the same group of proteins is consistently identified and precisely quantified across multiple samples, a requirement that is only partially achieved by current proteomics approaches. Selected reaction monitoring (SRM)—also called multiple reaction monitoring—is emerging as a technology that ideally complements the discovery capabilities of shotgun strategies by its unique potential for reliable quantification of analytes of low abundance in complex mixtures. In an SRM experiment, a predefined precursor ion and one of its fragments are selected by the two mass filters of a triple quadrupole instrument and monitored over time for precise quantification. A series of transitions (precursor/fragment ion pairs) in combination with the retention time of the targeted peptide can constitute a definitive assay. Typically, a large number of peptides are quantified during a single LC-MS experiment. This tutorial explains the application of SRM for quantitative proteomics, including the selection of proteotypic peptides and the optimization and validation of transitions. Furthermore, normalization and various factors affecting sensitivity and accuracy are discussed. |
format | Text |
id | pubmed-2583086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-25830862008-11-14 Selected reaction monitoring for quantitative proteomics: a tutorial Lange, Vinzenz Picotti, Paola Domon, Bruno Aebersold, Ruedi Mol Syst Biol Review Article Systems biology relies on data sets in which the same group of proteins is consistently identified and precisely quantified across multiple samples, a requirement that is only partially achieved by current proteomics approaches. Selected reaction monitoring (SRM)—also called multiple reaction monitoring—is emerging as a technology that ideally complements the discovery capabilities of shotgun strategies by its unique potential for reliable quantification of analytes of low abundance in complex mixtures. In an SRM experiment, a predefined precursor ion and one of its fragments are selected by the two mass filters of a triple quadrupole instrument and monitored over time for precise quantification. A series of transitions (precursor/fragment ion pairs) in combination with the retention time of the targeted peptide can constitute a definitive assay. Typically, a large number of peptides are quantified during a single LC-MS experiment. This tutorial explains the application of SRM for quantitative proteomics, including the selection of proteotypic peptides and the optimization and validation of transitions. Furthermore, normalization and various factors affecting sensitivity and accuracy are discussed. Nature Publishing Group 2008-10-14 /pmc/articles/PMC2583086/ /pubmed/18854821 http://dx.doi.org/10.1038/msb.2008.61 Text en Copyright © 2008, EMBO and Nature Publishing Group http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits distribution and reproduction in any medium, provided the original author and source are credited. Creation of derivative works is permitted but the resulting work may be distributed only under the same or similar licence to this one. This licence does not permit commercial exploitation without specific permission. |
spellingShingle | Review Article Lange, Vinzenz Picotti, Paola Domon, Bruno Aebersold, Ruedi Selected reaction monitoring for quantitative proteomics: a tutorial |
title | Selected reaction monitoring for quantitative proteomics: a tutorial |
title_full | Selected reaction monitoring for quantitative proteomics: a tutorial |
title_fullStr | Selected reaction monitoring for quantitative proteomics: a tutorial |
title_full_unstemmed | Selected reaction monitoring for quantitative proteomics: a tutorial |
title_short | Selected reaction monitoring for quantitative proteomics: a tutorial |
title_sort | selected reaction monitoring for quantitative proteomics: a tutorial |
topic | Review Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2583086/ https://www.ncbi.nlm.nih.gov/pubmed/18854821 http://dx.doi.org/10.1038/msb.2008.61 |
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