Measure intracelluar ROS - Protocol for DCF-DA staining? (Jan/08/2009 )

Hi All,

I would like to use DCF-DA to measure intracellular ROS by flow cytometry. Many people have published results using this method, but I haven't been able to find a detailed protocol using this method. Does anyone have a protocol they could share?

Also, suggestions on other assays to measure ROS production are needed and most welcomed!

Thanks!

Hi all,

Be careful if you use trypsine to dettach your cells. Trypsinization increase the ROS levels on the cells. We measure ROS by flow cytometry by H2DCF-DA (Molecular Probes) and results for cells in suspension are fine, but for other cell lines (endometrial adenocarcinoma) cells showed high levels of intracellular ROS after trypsinization