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Measuring ROS with DCFH probe

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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Measuring ROS with DCFH probe

Postby rem » Wed Oct 30, 2013 6:49 pm

Hello everyone,
This is my first posting on this forum and I would really appreciate your opinions on the folllowing situation.
I'm trying to measure ROS in HUVEC cells with the fluorescent probe DCFH by flow cytometry. I apply oxidant and antioxidant treatments, and what I get is that the cells with an oxidant treatment show less fluorescence than the control and those with oxidant + antioxidant. The curious thing is that the fluorescence pattern I see is exactly opposite to what I expect (I'm doing concentration curves).
I've repeated the experiment and I continue to get the same results. I'm sure that the treatments were applied correctly. Could this be an issue of the reading at the flow cytometer (setting, etc)?
Thanks in advance!
rem
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Postby JackBean » Wed Mar 05, 2014 10:20 am

What is the probe supposed to detect? The ROS directly?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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