Organoid Sample Preparation and Extraction for LC-MS Peptidomics
Summary: This protocol describes the peptidomic analysis of organoid lysates, FACS-purified cell populations, and 2D culture secretions by liquid chromatography mass spectrometry (LC-MS). Currently, most peptides are quantified by ELISA, limiting the peptides that can be studied. However, an LC-MS-b...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720301519 |
| _version_ | 1818921362067030016 |
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| author | Emily L. Miedzybrodzka Rachel E. Foreman Sam G. Galvin Pierre Larraufie Amy L. George Deborah A. Goldspink Frank Reimann Fiona M. Gribble Richard G. Kay |
| author_facet | Emily L. Miedzybrodzka Rachel E. Foreman Sam G. Galvin Pierre Larraufie Amy L. George Deborah A. Goldspink Frank Reimann Fiona M. Gribble Richard G. Kay |
| author_sort | Emily L. Miedzybrodzka |
| collection | DOAJ |
| description | Summary: This protocol describes the peptidomic analysis of organoid lysates, FACS-purified cell populations, and 2D culture secretions by liquid chromatography mass spectrometry (LC-MS). Currently, most peptides are quantified by ELISA, limiting the peptides that can be studied. However, an LC-MS-based approach allows more peptides to be monitored. Our group has previously used LC-MS for tissue peptidomics and secretion of enteroendocrine peptides from primary culture. Now, we extend the use to organoid models.For complete details on the use and execution of this protocol, please refer to Goldspink et al. (2020). |
| first_indexed | 2024-12-20T01:36:26Z |
| format | Article |
| id | doaj.art-a142a25934e846f4b27a6c18f2bc79b4 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-20T01:36:26Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a142a25934e846f4b27a6c18f2bc79b42022-12-21T19:58:00ZengElsevierSTAR Protocols2666-16672020-12-0113100164Organoid Sample Preparation and Extraction for LC-MS PeptidomicsEmily L. Miedzybrodzka0Rachel E. Foreman1Sam G. Galvin2Pierre Larraufie3Amy L. George4Deborah A. Goldspink5Frank Reimann6Fiona M. Gribble7Richard G. Kay8Wellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UKWellcome Trust – MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UK; Corresponding authorSummary: This protocol describes the peptidomic analysis of organoid lysates, FACS-purified cell populations, and 2D culture secretions by liquid chromatography mass spectrometry (LC-MS). Currently, most peptides are quantified by ELISA, limiting the peptides that can be studied. However, an LC-MS-based approach allows more peptides to be monitored. Our group has previously used LC-MS for tissue peptidomics and secretion of enteroendocrine peptides from primary culture. Now, we extend the use to organoid models.For complete details on the use and execution of this protocol, please refer to Goldspink et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166720301519ProteomicsOrganoidsMass Spectrometry |
| spellingShingle | Emily L. Miedzybrodzka Rachel E. Foreman Sam G. Galvin Pierre Larraufie Amy L. George Deborah A. Goldspink Frank Reimann Fiona M. Gribble Richard G. Kay Organoid Sample Preparation and Extraction for LC-MS Peptidomics STAR Protocols Proteomics Organoids Mass Spectrometry |
| title | Organoid Sample Preparation and Extraction for LC-MS Peptidomics |
| title_full | Organoid Sample Preparation and Extraction for LC-MS Peptidomics |
| title_fullStr | Organoid Sample Preparation and Extraction for LC-MS Peptidomics |
| title_full_unstemmed | Organoid Sample Preparation and Extraction for LC-MS Peptidomics |
| title_short | Organoid Sample Preparation and Extraction for LC-MS Peptidomics |
| title_sort | organoid sample preparation and extraction for lc ms peptidomics |
| topic | Proteomics Organoids Mass Spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720301519 |
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