Robust neutrophil extracellular trap assay kit

This neutrophil extracellular trap (NET) assay kit enables the accurate quantification of extracellular NET-associated elastase activity. Unlike nucleic acid stains which can suffer from false positives due to DNA release from necrosis, this protocol includes a step to wash away non-NET associated elastase following NET generation. This offers a DNA-independent readout of NET generation by measuring absorbance of the specific cleaved elastase substrate (N-methoxysuccinyl-Ala-Ala-Pro-Val p-Nitroanilide) at 405nm. Both PMA and calcium ionophore A-23187 are included as stimuli for inducing NET generation.

Stimulated neutrophil with NETs and trapped Shigella Bacteria

Coloured scanning electron micrograph showing stimulated neutrophil with NETs and trapped Shigella bacteria. © Max Planck Institute for Infection Biology

Benefits of using this NET assay kit
• Non-dsDNA readout eliminating false positives from DNA release by necrosis
• Quantifiable results
• Strong signal
• Easy to follow protocol
• Elastase-based analysis
• Adaptable to multiple species by using enzymatic activity rather than NET component antibodies

Neutrophil extracellular trap (NET) assay kit standard curve

Material available for download
Neutrophil Extracellular Trap (NET) Assay Kit Manual
Neutrophil Extracellular Trap (NET) Assay Kit MSDS