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

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...

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Detalles Bibliográficos
Autores principales: Clement, Cristina C., Nanaware, Padma P., Stern, Lawrence J., Santambrogio, Laura
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264744/
https://www.ncbi.nlm.nih.gov/pubmed/34278334
http://dx.doi.org/10.1016/j.xpro.2021.100648
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author Clement, Cristina C.
Nanaware, Padma P.
Stern, Lawrence J.
Santambrogio, Laura
author_facet Clement, Cristina C.
Nanaware, Padma P.
Stern, Lawrence J.
Santambrogio, Laura
author_sort Clement, Cristina C.
collection PubMed
description 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).
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spelling pubmed-82647442021-07-16 A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis Clement, Cristina C. Nanaware, Padma P. Stern, Lawrence J. Santambrogio, Laura STAR Protoc Protocol 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). Elsevier 2021-07-06 /pmc/articles/PMC8264744/ /pubmed/34278334 http://dx.doi.org/10.1016/j.xpro.2021.100648 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Clement, Cristina C.
Nanaware, Padma P.
Stern, Lawrence J.
Santambrogio, Laura
A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title_full A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title_fullStr A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title_full_unstemmed A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title_short A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
title_sort protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264744/
https://www.ncbi.nlm.nih.gov/pubmed/34278334
http://dx.doi.org/10.1016/j.xpro.2021.100648
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