Need to know two things
1. What is the appropriate time frame to record RLUs? (i am using a microplate reader, Spectramax M2, can't set the recording time below 1min)
2. How will be the typical curve look like?
Need one standard curve pattern for ROS generation (in terms of RLUs over time period) to compare with my result.
I am not sure whether the results I am getting (1min interval for 30min) are correct or not.
Requesting your valuable suggestions and some examples.
Thanks in advance.
You need to understand NOX gene induction/expression by a stimulus - for this, do a time course experiment. From the time course experiment arbitrarily choose a time point where the expression is highest (both by protein and RNA).
When You do measurements make sure your media is buffered so cells can function normally without a pH shock, temp shock, to solve this problem the plate should not go beyond say 8-10 minutes total time outside the incubator.
Hope this help, good luck!
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