Mitochondrial ROS Detection Protocol

Mitochondrial ROS formation after treatment was assessed using the MitoSOX kit (ThermoFisher, Watlham, MA, USA) modified as described by Kauffman et al. [43 (link)]. First, 10⁵ cells were resuspended in 500 μL of PBS, and then 1 μL of a 2.5 μM MitoSOX dye solution in DMSO was added. After 20 min of incubation at 37 °C, cells were centrifuged and resuspended in 500 μL of fresh PBS. Subsequently, the cells were analysed by flow cytometry.

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Magacz M., Papież M., Kościelniak D., Jurczak A., Kędziora K., Pamuła E, & Krzyściak W. (2023). Safety Assessment of the Modified Lactoperoxidase System—In Vitro Studies on Human Gingival Fibroblasts. International Journal of Molecular Sciences, 24(3), 2640.

Publication 2023

MitoSOX kit

After treatment Cells Dmso Dye 2 Flow cytometry Mitochondrial formation Mitosox

Corresponding Organization : AGH University of Krakow

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Variable analysis

independent variables

Treatment

dependent variables

Mitochondrial ROS formation

control variables

Number of cells (10^5)
Volume of PBS (500 μL)
Concentration of MitoSOX dye (2.5 μM)
Incubation time (20 min)
Incubation temperature (37 °C)

controls

No controls were explicitly mentioned in the provided information.

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