ROS-Glo H2O2 Assay in MDA-MB-231 Cells

MDA-MB-231 cells were transferred to 96-well plates (20,000 cells/well) after treatment, with 60 μL fresh-media and were incubated for 12 h, as described previously^{20 (link),55 (link)}. At 7 h, 20 μL of H₂O₂ substrate solution from ROS-Glo H₂O₂ assay kit (Promega) was added to cells and incubated for 5 h more. At 12 h, cells were incubated for 20 min after addition of ROS-Glo™ detection solution and the Lum was measured at 0.5 s integration. Synergy LX Multi-Mode Reader was used for this purpose.

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Mittal L., Camarillo I.G., Varadarajan G.S., Srinivasan H., Aryal U.K, & Sundararajan R. (2020). High-throughput, Label-Free Quantitative Proteomic Studies of the Anticancer Effects of Electrical Pulses with Turmeric Silver Nanoparticles: an in vitro Model Study. Scientific Reports, 10, 7258.

Publication 2020

ROS-Glo H2O2 assay kit

Assay Cells H2o2 Mda mb 231 cells Promega

Corresponding Organization :

Other organizations : Purdue University West Lafayette, Anna University, Chennai, B.S. Abdur Rahman Crescent Institute of Science & Technology

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Variable analysis

independent variables

Treatment

dependent variables

Reactive oxygen species (ROS) production

control variables

Cell line (MDA-MB-231)
Cell density (20,000 cells/well)
Culture medium volume (60 μL)
Incubation time (12 h)
H2O2 substrate solution addition (20 μL at 7 h)
Incubation time after H2O2 substrate addition (5 h)
ROS-Glo™ detection solution addition (incubation for 20 min)
Luminescence measurement (0.5 s integration)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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