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Mitosox red

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MitoSOX Red is a fluorogenic dye that is highly selective for detecting superoxide in the mitochondria of live cells. It functions by rapidly entering live cells, where it is selectively targeted to the mitochondria and oxidized by superoxide. The oxidation product then exhibits red fluorescence, providing a reliable indicator of mitochondrial superoxide levels.

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Lab products found in correlation

Mitosox red, by Thermo Fisher Scientific (4 mentions) Calcium and magnesium, by Thermo Fisher Scientific (2 mentions) H2dcf da, by FlowJo (1 mentions) H2dcfda, by Beyotime (1 mentions) Facscanto 2, by BD (1 mentions)

4 protocols using mitosox red

1

Cellular Stress Evaluation by Flow Cytometry

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To assess the level of cellular stress, HConFs were inoculated into 6-well plates (4×104 cells-1). HConFs were treated with different treatments for a period of time and washed twice with PBS. The 500μl reaction solution at a concentration of 5x10-6m H2DCF-DA (Beyotime,1;1000) or MitoSOX Red (5μM, Invitrogen, M36008) for mitochondria-derived ROS evaluation) was incubated in the dark for 20 min at 37 °C. Next, HConFs were trypsinized and resuspended in 150μl of phosphate buffer solution (PBS). Finally, the oxidation of H2DCF-DA and MitoSOX Red were detected by flow cytometry and analyzed by Flow-Jo_V10 software.
Li X., Leng Y., Li X., Wang Y., Luo P., Zhang C., Wang Z., Yue X., Shen C., Chen L., Liu Z., Shi C, & Xie L. (2020). The TβR II-targeted aptamer S58 prevents fibrosis after glaucoma filtration surgery. Aging (Albany NY), 12(10), 8837-8857.
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2

Mitochondrial Superoxide Detection

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Cells were incubated for 30 minutes at 37°C with 0.5 μM MitoSOX Red (Thermo Fisher Scientific, Inc.). They were then collected by trypsinization, resuspended in 1X PBS and analyzed on a FACSCanto™II (BD Biosciences) instrument, before and after addition of Propidium Iodide. The appropriate negative (unstained) and positive (cells treated with 50 μM antimycin A for 1 h prior to the staining) controls were included. Analysis of MitoSOX Red fluorescence was performed with FlowJo™.
Papadaki V., Erpapazoglou Z., Kokkori M., Rogalska M.E., Potiri M., Birladeanu A., Tsakiri E.N., Ashktorab H., Smoot D.T., Papanikolopoulou K., Samiotaki M, & Kafasla P. (2023). IQGAP1 mediates the communication between the nucleus and the mitochondria via NDUFS4 alternative splicing. NAR Cancer, 5(3), zcad046.
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3

Quantifying Mitochondrial Superoxide

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Fibroblasts were plated in a 6 well plate at 300,000 cells per well. After incubation for 24 hours, cells were loaded with 1ml of 5 μM MitoSOX Red (Invitrogen) and incubated for 10 minutes at 37°C in 5% CO2 atmosphere. Cells were washed three times with Hank’s balanced salt solution with calcium and magnesium (Gibco), trypsinized and placed in alpha MEM. Using a flow cytometer (FlowJo, LLC), MitoSOX Red was excited at 488 nm and fluorescence emission at 575 was measured. Fluorescense was quantified using fluorescence activated cell sorting (FACS).
Abraham M., Collins C.A., Flewelling S., Camazine M., Cahill A., Cade W.T, & Duncan J.G. (2018). Mitochondrial inefficiency in infants born to overweight African American mothers. International journal of obesity (2005), 42(7), 1306-1316.
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4

Quantifying Mitochondrial Superoxide

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Fibroblasts were plated in a 6 well plate at 300,000 cells per well. After incubation for 24 hours, cells were loaded with 1ml of 5 μM MitoSOX Red (Invitrogen) and incubated for 10 minutes at 37°C in 5% CO2 atmosphere. Cells were washed three times with Hank’s balanced salt solution with calcium and magnesium (Gibco), trypsinized and placed in alpha MEM. Using a flow cytometer (FlowJo, LLC), MitoSOX Red was excited at 488 nm and fluorescence emission at 575 was measured. Fluorescense was quantified using fluorescence activated cell sorting (FACS).
Abraham M., Collins C.A., Flewelling S., Camazine M., Cahill A., Cade W.T, & Duncan J.G. (2018). Mitochondrial inefficiency in infants born to overweight African American mothers. International journal of obesity (2005), 42(7), 1306-1316.
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