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

Cristina C. Clement; Padma P. Nanaware; Lawrence J. Stern; Laura Santambrogio

STAR Protocols (Sep 2021)

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

Cristina C. Clement,
Padma P. Nanaware,
Lawrence J. Stern,
Laura Santambrogio

Affiliations

Cristina C. Clement: Department of Radiation Oncology, Weill Cornell Medicine, New York, NY 10065 USA; Corresponding author
Padma P. Nanaware: Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA
Lawrence J. Stern: Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA
Laura Santambrogio: Department of Radiation Oncology, Weill Cornell Medicine, New York, NY 10065 USA; Caryl and Israel Englander Institute for Precision Medicine, New York, NY 10065, USA; Sandra and Edward Meyer Cancer Center, New York, NY 10065, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 3
p. 100648

Abstract

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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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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