UMMS Affiliation

Department of Pathology

Publication Date

2021-07-06

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Biological Factors | Investigative Techniques

Abstract

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).

Keywords

antigen processing and presentation, endosomes, MHC II

Rights and Permissions

Copyright 2021 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

DOI of Published Version

10.1016/j.xpro.2021.100648

Source

Clement CC, Nanaware PP, Stern LJ, Santambrogio L. A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis. STAR Protoc. 2021 Jul 6;2(3):100648. doi: 10.1016/j.xpro.2021.100648. PMID: 34278334; PMCID: PMC8264744. Link to article on publisher's site

Journal/Book/Conference Title

STAR protocols

Related Resources

Link to Article in PubMed

PubMed ID

34278334

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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