CM-H2DCFDA (General Oxidative Stress Indicator)

Protocol tips

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	- Cells were treated with 5 µM CM-H2DCFDA. - Cells were incubated at 37C for 20 min.

Protocol tips
- Cells were treated with 5 µM CM-H2DCFDA. - Cells were incubated at 37C for 20 min.

Publication protocol

Cells were seeded at 80% confluency in a 60 mm culture dishes a day prior to drug treatment or transfection. To measure intracellular ROS level, cell pellets were re-suspended in RPMI-1640 medium containing 5 µM of 5-(and-6)-chloromethyl-2-,7-dichlorofluorescin diacetate (CM-H2DCFDA; Molecular Probes, Life Technologies) and incubated in the dark at 37 °C for 20 minutes. Cell suspensions were further diluted and filtered for flow cytometric analysis using BD LSRFortessaTM cell analyzer (BD Biosciences) with excitation and emission spectra of 495 nm and 529 nm respectively.

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