To evaluate intracellular ROS level, U251 cells were incubated using 10 μM DCFH-DA (Solarbio, CA1410) for 20 min at 37°C after BCA treatment, then washed with serum-free medium three times and imaged under Olympus fluorescence microscope (BX53). Meanwhile, the cell was digested with trypsin and resuspended. The ROS level was analyzed by flow cytometry (BD FACSCanto™ low cytometry, USA).
Facscanto low cytometry
Manufactured by BD
Sourced in United States
The BD FACSCanto™ low cytometer is a flow cytometry instrument designed for research applications. It provides multi-parameter analysis of cells and other particles in a fluid sample.
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Lab products found in correlation
3 protocols using facscanto low cytometry
1
Quantifying Intracellular ROS in U251 Cells
2
Apoptosis Assay of U251 Cells
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U251 cells were treated with 0, 50, and 100 μmol/L BCA for 48 h. Then, U251 cells (1 × 106) were collected, after which 5 µL of PE Annexin V and 5 µL of 7-AAD were added. Gently vortex the cells and incubate at 25 °C in the dark for 15 min and the suspension was analyzed by flow cytometry (BD FACSCanto™ low cytometry, USA).
3
Apoptosis Analysis of U251 Cells
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The percentage of apoptotic cells was tested by Annexin V-PE/FITC (BD, Biosciences). U251 cells were treated with 0, 50, and 100 μmol/L BCA for 48 h. Then, U251 cells (1 × 106) were collected, after which 5 µl of PE Annexin V and 5 µl of 7-AAD were added. The cells were gently vortexed at room temperature and incubated for 15 minutes in the dark and the suspension was analyzed by flow cytometry (BD FACSCanto™ low cytometry, USA).
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