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. 2021 Jan 5;11(1):e3877. doi: 10.21769/BioProtoc.3877

Figure 3. Protocol schematic.

Figure 3.

Cells are loaded with DCF-based probe, incubated for desired treatment period (e.g., 1 h or 24 h) before plate spectroscopy readout of DCF as an indication of ROS generation. To normalize DCF signal to cell population per well, a simultaneous acidic protein precipitation stage and cellular protein dye stage is performed before solubilization and spectroscopic readout of SRB. The TCA-SRB assay can be utilized to obtain relative cell populations for growth inhibition, cytotoxicity or to normalize other cell-based assay readouts.