We are looking to do a time dependent mtROS measurement after MPP+ treatment given to cells. Can anyone suggest if pretreatment with MitoSOX ( followed by washing) could be given before cells are treated with MPP+. I have found papers following this procedure for the total ROS estimation using DCFDA but couldn't find using MitoSOX.
ROS is measured by nonfluorescent H2DCFDA to DCF, but DCF is easily diffused from the cells as well as H2DCFDA may interact with another compound. In this case, washing H2DCFDA is needed. MitoSox is incorporated into mitochondria where they react with ROS. The following paper uses mitoSOX prewashed and used for their assay:
Article Sophora flavescens Aiton Decreases MPP+-Induced Mitochondria...
The original MitoSOX and other dyes are described below. The company has been changed a few times from molecular probe to invitrogen to fisherthermo. If they still have people from the original company (molecular probe) working there, they will be able to answer your tech questions:
https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook/probes-for-reactive-oxygen-species-including-nitric-oxide/generating-and-detecting-reactive-oxygen-species.html
Thank you for your reply. However, the paper listed here has given treatment with MitoSox after 24h of MPP+ treatment.
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