biostudies-literatureUnknown23(11)Ghosh SNeutrophils are indispensable for defense against pathogens. Injured tissue-infiltrated neutrophils can establish a niche of chronic inflammation and promote degeneration. Studies investigated transcriptome of single-infiltrated neutrophils which could misinterpret molecular states of these post mitotic cells. However, neutrophil proteome characterization has been challenging due to low harvests from affected tissues. Here, we present a workflow to obtain proteome of 1000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6200 mouse and ∼5300 human proteins from circulating neutrophils. 4800 mouse and 3400 human proteins were recovered from 1000 cells with 10²-10⁸ copies/cell. Neutrophils from stroke-affected mouse brains adapMolecular & cellular proteomics : MCP100858https://www.ebi.ac.uk/biostudies/studies/S-EPMC11630641biostudies-literatureProteomic Characterization of 1000 Human and Murine Neutrophils Freshly Isolated From Blood and Sites of Sterile Inflammation.PMC11630641Lange EGruneboom ARichter MSickmann AHeyer RStenzel MSoehnlein OBeer AGhosh STuz AANagel DCibir ZJung MHermann DMHasenberg AGunzer MSingh VfalseProteomic Characterization of 1000 Human and Murine Neutrophils Freshly Isolated From Blood and Sites of Sterile Inflammation.Neutrophils are indispensable for defense against pathogens. Injured tissue-infiltrated neutrophils can establish a niche of chronic inflammation and promote degeneration. Studies investigated transcriptome of single-infiltrated neutrophils which could misinterpret molecular states of these post mitotic cells. However, neutrophil proteome characterization has been challenging due to low harvests from affected tissues. Here, we present a workflow to obtain proteome of 1000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6200 mouse and ∼5300 human proteins from circulating neutrophils. 4800 mouse and 3400 human proteins were recovered from 1000 cells with 10²-10⁸ copies/cell. Neutrophils from stroke-affected mouse brains adap2024-01-01T00:00:00Z2024 Nov2026-06-02T04:38:12.765Z2025-04-19T14:35:59.206ZS-EPMC116306413939558110.1016/j.mcpro.2024.100858