Selected reaction monitoring for quantitative proteomics: a tutorial
Abstract 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 reacti...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2008-10-01
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| Series: | Molecular Systems Biology |
| Online Access: | https://doi.org/10.1038/msb.2008.61 |
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| _version_ | 1849225777422270464 |
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| author | Vinzenz Lange Paola Picotti Bruno Domon Ruedi Aebersold |
| author_facet | Vinzenz Lange Paola Picotti Bruno Domon Ruedi Aebersold |
| author_sort | Vinzenz Lange |
| collection | DOAJ |
| description | Abstract 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 | Article |
| id | doaj-art-8d79fb43a6c742f28dedeb285b0b9fb3 |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2008-10-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-8d79fb43a6c742f28dedeb285b0b9fb32025-08-24T12:01:33ZengSpringer NatureMolecular Systems Biology1744-42922008-10-014111410.1038/msb.2008.61Selected reaction monitoring for quantitative proteomics: a tutorialVinzenz Lange0Paola Picotti1Bruno Domon2Ruedi Aebersold3Institute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichAbstract 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.https://doi.org/10.1038/msb.2008.61 |
| spellingShingle | Vinzenz Lange Paola Picotti Bruno Domon Ruedi Aebersold Selected reaction monitoring for quantitative proteomics: a tutorial Molecular Systems Biology |
| 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 |
| url | https://doi.org/10.1038/msb.2008.61 |
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