- Organoid Sample Preparation and Extraction for LC-MS Peptidomics.
Organoid Sample Preparation and Extraction for LC-MS Peptidomics.
STAR protocols (2020-12-31)
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
PMID33377058
摘要
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).
材料
产品编号
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产品描述
Sigma-Aldrich
D -(+)-葡萄糖, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
白蛋白 溶液 来源于牛血清, 10% in DPBS, low endotoxin, fatty acid free, suitable for cell culture, sterile-filtered
Sigma-Aldrich
汉克斯平衡盐 溶液, Modified, with sodium bicarbonate, without calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture