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

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

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...

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Bibliographic Details
Main Authors: Cristina C. Clement, Padma P. Nanaware, Lawrence J. Stern, Laura Santambrogio
Format: Article
Language:English
Published: Elsevier 2021-09-01
Series:STAR Protocols
Subjects:
cell biology
cell separation/fractionation
immunology
mass spectrometry
protein biochemistry
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721003555
Description
Summary: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).
ISSN:2666-1667

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