A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis

STAR Protocols

Padma P. Nanaware¹  Lawrence J. Stern²  Laura Santambrogio²  Cristina C. Clement³ 

[1] Corresponding author;Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA;Department of Radiation Oncology, Weill Cornell Medicine, New York, NY 10065 USA;

关键词: cell biology;    cell separation/fractionation;    immunology;    mass spectrometry;    protein biochemistry;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown