1,3-Diphenylisobenzofuran (solution)

1,3-Diphenylisobenzofuran (DPBF) (solution) has been developed as a selective probe for the detection and quantitative determination of hydrogen peroxide in samples containing different reactive nitrogen and oxygen species (RNOS). DPBF is a fluorescent probe which, for almost 20 years, was believed to react in a highly specific manner toward some reactive oxygen species such as singlet oxygen and hydroxy, alkyloxy or alkylperoxy radicals^[1].
Solvent and concentration: DMSO: 10 mM
The 1 mL volume is defined as the base specification. All larger sizes correspond to incremental volumes of this base. In Vitro:General Protocol

Preparation of DPBF working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of DPBF in 3.6992 mL of DMSO to obtain 10 mM of DPBF.
Note: It is recommended to store the stock solution at -20°C, -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of DPBF working solution
Dilute the stock solution in serum-free cell culture medium or PBS. 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: Please adjust the concentration of DPBF working solution according to the actual situation.

Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of DPBF working solution, and then incubate at room temperature for 30 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

Precautions
1. It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles.
2. Please adjust the concentration of DPBF working solution according to the actual situation.
3. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.