Neutrophils were stimulated for 4 hours with 100 nM PMA (Sigma-Aldrich), and NET formation was verified by staining with sytoxgreen (125 nM, Thermo Fisher Scientific). The cell culture medium was aspirated, cells were washed three times with PBS, and NETs were detached by scraping, sonicated, and quantified by PicoGreen assay (Thermo Fisher Scientific). Either DNA or proteins from NETs were isolated by phenol-chloroform extraction, or NETs were digested. The resulting fractions were run on an agarose gel to test the DNA content. To induce degranulation, neutrophils were stimulated with fMLP (10 μM, Sigma-Aldrich).

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.