I am currently setting up a two step treatment on HepG2 cells in a 96 well plate, using an antioxidant and a drug. Majority of the literature that I have consulted mention a pretreatment with the antioxidant, and then treatment using a drug. For the few treatments that I have set up, I give the tretament using antioxidant dissolved in media for 24 hours, remove the media, wash with PBS and then give the drug tretament (drug dissolved in media). My confusion here is : Is this protocol correct? Or do I keep the antioxidant containing media and just add the drug containing media on top of the antioxidant treatment, without removing the antioxidant and forgoing the PBS wash. Any clarification would be appreciated
Dear Malhar Desai,
Your using protocol is correct. But you do must without PBS washing. Both set up give best results.
Thank you
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