MitoPerOx (solution)

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Solution preparation^[2]
1.1 Stock solution
Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute with PBS or cell culture medium (optimized according to the experiment). The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: The working solution should be prepared and used immediately. Keep it away from light.
2. MitoPerOx Assay Protocol:
2.1 Cells (e.g., HEK293 or fibroblasts) are plated on coverslips in 35 mm dishes and cultured at 37 °C, 5% CO2.
2.2 Probe Preparation: Add 100 nM MitoPerOx to phenol red-free medium and incubate at 37 °C for 30 min in the dark.
2.3 Washing: Discard medium and wash cells three times with PBS.
2.4 Induce oxidative stress: Treat cells with 500 μM H₂O₂ in Hepes-Tris buffer for 15-30 min.
2.5 Fluorescence imaging: Confocal microscopy (excitation wavelength 488 nm) was used to collect emission light at 520 nm (oxidized state) and 590 nm (reduced state) , and the extent of lipid peroxidation was quantified by calculating the 520/590 nm ratio.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.