Organoid Sample Preparation and Extraction for LC-MS Peptidomics

被引：4

作者：

Miedzybrodzka, Emily L. ^{[1
]}

Foreman, Rachel E. ^{[1
]}

Galvin, Sam G. ^{[1
]}

Larraufie, Pierre ^{[1
,2
]}

George, Amy L. ^{[1
]}

Goldspink, Deborah A. ^{[1
,3
]}

Reimann, Frank ^{[1
]}

Kay, Richard G. ^{[1
]}

机构：

[1] Univ Cambridge, Addenbrookes Hosp, Inst Metab Sci Metab Res Labs, Wellcome Trust MRC, Hills Rd, Cambridge, England

[2] Univ Paris Saclay, AgroParisTech, INRAE, UMR PNCA, F-75005 Paris, France

[3] GSK Med Res Ctr, Novel Human Genet, Stevenage, England

来源：

STAR PROTOCOLS | 2020年 / 1卷 / 03期

基金：

英国惠康基金;

关键词：

Mass Spectrometry; Organoids; Proteomics;

D O I：

10.1016/j.xpro.2020.100164

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This protocol describes the peptidomic analysis of organoid lysates, FACS-puri-fied 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).

引用

页数：19

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