Hydrogen peroxide cell based assay kit

Cells were incubated for 2 h with either compounds (30 μM) or DMSO in a 96-well plate. After the treatment, 80 μl of cell culture medium was collected and the level of hydrogen peroxide was determined using a Hydrogen Peroxide Cell-Based Assay kit (Cayman).

The extracellular H₂O₂ produced by TNBC cells was measured using a hydrogen peroxide cell-based assay kit (Cayman Chemical, Arbor, MI, USA). Using horseradish peroxidase (HRP) as a catalyst, this extremely sensitive and stable H₂O₂ probe interacts with extracellular H₂O₂ to create the highly fluorescent resorufin. Briefly, cells were plated at a density of 5 × 10⁴ per 100 µL in 96-well plates in a serum-free culture medium. The cells were exposed to TQ at different concentrations (5, 10, and 15 µM) for 24 h after overnight incubation. For the treatment controls and catalase controls, extra wells for media and working solutions without cells were allotted, as instructed in the kit’s instruction booklet. According to the directions in the kit, the samples were examined. Using a microplate reader, the fluorescence intensity of each well (530 nm for excitation and 590 nm for emission) was measured.