[HTML][HTML] Immunodetection of NETs in paraffin-embedded tissue

V Brinkmann, U Abu Abed, C Goosmann… - Frontiers in …, 2016 - frontiersin.org
V Brinkmann, U Abu Abed, C Goosmann, A Zychlinsky
Frontiers in immunology, 2016frontiersin.org
The pathogenic potential of neutrophil extracellular traps (NETs) was recently described,
and their detection in tissue could serve as a prognostic marker. NETs are delicate and
filigree structures; hence good tissue preservation is essential for their detection. Indeed,
analysis of paraffin-embedded tissue has proven superior to the study of cryo sections.
Though, under favorable conditions, the presence of NETs can be detected in tissue
sections stained with histological dyes, definitive identification of NETs needs the …
The pathogenic potential of neutrophil extracellular traps (NETs) was recently described, and their detection in tissue could serve as a prognostic marker. NETs are delicate and filigree structures; hence good tissue preservation is essential for their detection. Indeed, analysis of paraffin-embedded tissue has proven superior to the study of cryo sections. Though, under favorable conditions, the presence of NETs can be detected in tissue sections stained with histological dyes, definitive identification of NETs needs the colocalization of immunofluorescent signals for both nuclear and granular (or cytoplasmic) NET components. We tested diverse antigen retrieval methods and various combinations of commercially available antibodies and present here staining protocols to detect NETs in human and murine tissue sections.
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