Organoid Sample Preparation and Extraction for LC-MS Peptidomics.
Organoid Sample Preparation and Extraction for LC-MS Peptidomics.
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DOI：
10.1016/j.xpro.2020.100164
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发表时间：
2020-12-18
期刊：
影响因子：
--
通讯作者：
Kay RG
中科院分区：
文献类型：
--
作者：
Miedzybrodzka EL;Foreman RE;Galvin SG;Larraufie P;George AL;Goldspink DA;Reimann F;Gribble FM;Kay RG
该方案描述了通过液相色谱质谱法（LC-MS）对类器官裂解物、FACS纯化的细胞群和2D培养物分泌物进行的肽组分析。目前，大多数肽通过ELISA定量，限制了可以研究的肽。然而，基于LC-MS的方法允许监测更多的肽。我们的小组以前使用LC-MS的组织肽组学和分泌的肠内分泌肽从原代培养。现在，我们将其应用扩展到类器官模型。有关本方案使用和执行的完整详细信息，请参阅。制备用于肽组学的类器官培养物、分泌物和FAC-sorted细胞已建立的管道，包括肽提取、LC-MS和数据分析密切相关的肽激素的非环境生物学检测来自刺激的类器官培养物的分泌肽的半定量分析该方案描述了类器官裂解物、FACS纯化的细胞群、和通过液相色谱质谱法（LC-MS）的2D培养物分泌物。目前，大多数肽通过ELISA定量，限制了可以研究的肽。然而，基于LC-MS的方法可以监测更多的肽。我们的小组以前使用LC-MS的组织肽组学和分泌的肠内分泌肽从原代培养。现在，我们将其应用扩展到类器官模型。
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. Preparing organoid cultures, secretions, and FAC-sorted cells for peptidomics An established pipeline covering peptide extraction, LC-MS, and data analysis Unambiguous detection of closely related peptide hormones Semi-quantitative analysis of secreted peptides from stimulated organoid cultures 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.
影响因子：
64.5
作者：
Beumer, Joep;Puschhof, Jens;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
8.8
作者：
Goldspink, Deborah A.;Lu, Van B.;Gribble, Fiona M.
通讯作者：
Gribble, Fiona M.
影响因子：
14.8
作者：
Fujii, Masayuki;Matano, Mami;Sato, Toshiro
通讯作者：
Sato, Toshiro