Simplified flow cytometric quantification of human neutrophil extracellular traps (NETs).

ABSTRACT: Several cellular pathways lead to the formation of neutrophil extracellular traps (NETs), a form of cell death (NETosis), distinct from apoptotic and necrotic cell death. Surprisingly, there remains a paucity of methods enabling efficient quantification of NETosis and associated pathways. Here, we describe the development of a simple, sensitive, reliable and flexible flow cytometry assay allowing efficient detection and quantification of NETosis. For the core assay, isolated primary human neutrophils were incubated with stimulants e.g. PMA/ionomycin with or without inhibitors prior to fixing. The fixed cells were then blocked and subsequently incubated in anti-DNA/Histone 1 and anti-histone H2A antibodies for dual detection of Histone 1 and Histone H2A. Cells with were H1-DNA/H2A double fl

SUBMITTER: Rinaldi G 

PROVIDER: S-EPMC12947343 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature