A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis
UMass Chan AffiliationsDepartment of Pathology
Keywordsantigen processing and presentation
Amino Acids, Peptides, and Proteins
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AbstractA 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).
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