I have taken H2DCFDA fluoresce at 485/535 and now I want make a ROS relative production bar graph for values. Kindly suggest different steps.
Hey Sandeep,
although I favorize raw data (also for ROS), you can calculate the relative ROS levels with these steps:
1. Substract your medium controls without cells (with or without any substances or treatment) from the according groups.
2. Define your untreated group (for example cells, which only got normal medium without substances or any kind of treatment).
3. Divide all your other values by your control group (this will deliver you a zero line for the control group and relative values for all other groups).
I hope that helps you.
All the best,
Marc
In context of this topic, I would like to bring to your attention a recent review of our group, "Functions of ROS in macrophages and antimicrobial immunity".
In this review, we give an introduction to ROS and their sources in macrophages, summarize the versatile roles of ROS in direct and indirect antimicrobial immune defense and provide an overview of commonly used ROS probes, ROS source inhibitors and ROS scavengers (also the difference between ROS scavengers and antioxidants, which are not synonymous, is explained).
If you like, please have a look at:
Functions of ROS in Macrophages and Antimicrobial Immunity
February 2021, Antioxidants 10(2):313
DOI: https://doi.org/10.3390/antiox10020313
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