| Title | A protocol for qualitative and quantitative measurement of endosomal processing using hot spot analysis. |
| Publication Type | Journal Article |
| Year of Publication | 2021 |
| Authors | Clement CC, Nanaware PP, Stern LJ, Santambrogio L |
| Journal | STAR Protoc |
| Volume | 2 |
| Issue | 3 |
| Pagination | 100648 |
| Date Published | 2021 Sep 17 |
| ISSN | 2666-1667 |
| Keywords | Animals, Antigens, Dendritic Cells, Endosomes, Membrane Proteins, Mice, Molecular Biology, Peptides, Tandem Mass Spectrometry |
| 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). |
| DOI | 10.1016/j.xpro.2021.100648 |
| Alternate Journal | STAR Protoc |
| PubMed ID | 34278334 |
| PubMed Central ID | PMC8264744 |
| Grant List | P01 AG031782 / AG / NIA NIH HHS / United States |