Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
- 50,000 cells/well were seeded |
- Cells werre treated with α-TOS, NTα-TOS-NPs, or T-α-TOS-NPs for 12 h at 37 °C in 5% CO2.
- RIPA cell lysis buffer was
added and mixed for 2 min |
|
Upstream tips |
- 50,000 cells/well were seeded |
Protocol tips |
- Cells werre treated with α-TOS, NTα-TOS-NPs, or T-α-TOS-NPs for 12 h at 37 °C in 5% CO2.
- RIPA cell lysis buffer was
added and mixed for 2 min |
Publication protocol
Intracellular ROS was assessed by the ROS assay Kit from Cell Biolabs. MCF-7 and H9C2 cells (50,000 cells/well) were plated in 96-well plates in 100 µL medium and grown for overnight. Then the medium was replenished and cells were treated with α-TOS, NTα-TOS-NPs, or T-α-TOS-NPs (40 µM for MCF-7, 60 µM for H9C2 with respect to α-TOS) for 12 h at 37 °C in 5% CO2 atmosphere. As a control, oligomycin (25 µM) treatment was performed. The media was removed and 100 µL of 1X dichloro-dihydro-fluorescein diacetate (DCFH-DA)/media solution was added to the cells and incubated for 45 min. The solution was removed and washed with PBS twice. Finally, RIPA cell lysis buffer was added and mixed for 2 min. The fluorescence was recorded at 485/530 nm.
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