Clement, Cristina C.Nanaware, Padma P.Stern, Lawrence J.Santambrogio, Laura2022-08-232022-08-232021-07-062022-02-03<p>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. <a href="https://doi.org/10.1016/j.xpro.2021.100648">Link to article on publisher's site</a></p>2666-1667 (Linking)10.1016/j.xpro.2021.10064834278334https://hdl.handle.net/20.500.14038/42063A 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).en-USCopyright 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/).http://creativecommons.org/licenses/by-nc-nd/4.0/antigen processing and presentationendosomesMHC IIAmino Acids, Peptides, and ProteinsBiological FactorsInvestigative TechniquesJournal Articlehttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=5898&amp;context=oapubs&amp;unstamped=1https://escholarship.umassmed.edu/oapubs/486527978615oapubs/4865