Wellcome Trust - MRC Institute of Metabolic Science Metabolic Research Laboratories, University of Cambridge, Addenbrookes Hospital, Hills Road, Cambridge, UK.
STAR Protoc. 2020 Nov 25;1(3):100164. doi: 10.1016/j.xpro.2020.100164. eCollection 2020 Dec 18.
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).
本方案描述了通过液相色谱-质谱联用（LC-MS）对类器官裂解物、FACS 纯化细胞群和 2D 培养物分泌液进行肽组学分析。目前，大多数肽类物质都是通过 ELISA 进行定量检测的，这限制了可以研究的肽类物质的种类。然而，基于 LC-MS 的方法可以监测更多的肽类物质。我们小组之前曾使用 LC-MS 进行组织肽组学研究和原代培养的肠内分泌肽的分泌研究。现在，我们将其应用于类器官模型。有关此方案使用和执行的完整详细信息，请参考 Goldspink 等人（2020 年）的研究。
