使用热点分析定量和定性测量内体处理的方案。

Department of Radiation Oncology, Weill Cornell Medicine, New York, NY 10065 USA.

Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA.

STAR Protoc. 2021 Jul 6;2(3):100648. doi: 10.1016/j.xpro.2021.100648. eCollection 2021 Sep 17.

A detailed quantification of antigen processing by endosomal compartments provides important information on the pattern of protein fragmentation. Here, we describe a protocol that combines gradient purified endosomes, incubated with antigens, followed by hot spot analysis of MS/MS-sequenced peptides. The analysis identifies differences in endosomal antigen processing by dendritic cells under diverse experimental conditions. For complete details on the use and execution of this protocol, please refer to Clement et al. (2021).

详细量化内体区室中的抗原加工可提供关于蛋白质片段化模式的重要信息。在这里，我们描述了一种结合梯度纯化内体与抗原孵育，然后对 MS/MS 测序肽进行热点分析的方案。该分析可识别在不同实验条件下树突状细胞中内体抗原加工的差异。有关该方案的使用和执行的完整详细信息，请参阅 Clement 等人。（2021 年）。