DCFDA / H2DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 25000 cells per well were platted in 96 well plates in DMEM without phenol red	- After 24h cells were treated with 25 μM DCFDA and incubated for 45 min at 37°C. - For 10 min to 1 h they were treated with 0, 500 or 1000 μM H2O2.

Upstream tips
- 25000 cells per well were platted in 96 well plates in DMEM without phenol red
Protocol tips
- After 24h cells were treated with 25 μM DCFDA and incubated for 45 min at 37°C. - For 10 min to 1 h they were treated with 0, 500 or 1000 μM H2O2.

Publication protocol

Intracellular reactive oxygen species were measured with the cell-permeant fluorogenic dye 2’,7’-dichlorofluorescein diacetate (DCFDA) as recommended by the manufacturer. Myoblasts were plated at a density of 25000 cells per well in 96 well plates in DMEM without phenol red. The following day cells were labeled with DCFDA (25 μM) 45 min at 37°C and treated with 0, 500 or 1000 μM H2O2 for 10 min to 1 h. Fluorescence intensity was measured with a fluorescence plate reader (TRIAD LT detector from Dynex; λexc = 485 nm; λem = 535 nm). Each condition was done in triplicate.

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Manufacturer protocol

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