Organoid Sample Preparation and Extraction for LC-MS Peptidomics

用于 LC-MS 肽组学的类器官样品制备和提取

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作者: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

Abstract

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).

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