Because cells I'm using express GFP, I use CellRox for ROS detection. Unfortunately, I'm not able to see differences between my conditions as expected. I need to know if CellRox could detect ROS variations in my model and I'm looking for a positive control of intracellular ROS generation. I tried with H202 but without concluant results.
Thanks.
Hi Colette
Which CellRox are you using? If it's CellRox Red than it's not supposed to interfere with your assay. However CellRox Green will interfere with GFP since the have similar spectra.
As for a positive control, H2O2 is the better one. What concentration and incubation period of H2O2 are you using?
What concentration of the H2O2 did you use? 800uM should do. Incubation time should not make that much of a difference as the H2O2 will be quickly converted within the cells to ROSs. I have tried it and it works very nicely, also, as mentioned above, I used the cell rox that uses the APC channel for detection (Cellrox Orange). Good luck
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