In vitro oxidation assay was performed as described previously (18). Briefly, a reaction (100 μl total volume) containing 5 μg of substrates (purified RIG-I, KSHV helicase, or MCM7), 2 μg of enzymes (purified viperin or viperin-Del), 50 mM Hepes (pH 7.5), 150 mM KCl, 2 mM SAM, and 5 mM dithionite was incubated for 45 min at 37°C. Reaction was stopped by boil for 10 min. Reactions containing RIG-I, MCM7, and KSHV helicase were analyzed by mass spectrometry, and the ratio of oxidized peptides over total peptides was quantitatively determined.

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.