1,3-Diphenylisobenzofuran  (Synonyms: DPBF)

1,3-Diphenylisobenzofuran (DPBF) 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].

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
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 to obtain 20 μM of DPBF working solution.
Note: Please adjust the concentration of DPBF working solution according to the actual situation.
2. 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.
3. Precautions
3.1 It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles.
3.2 Please adjust the concentration of DPBF working solution according to the actual situation.
3.3 This product is for R&D use only, not for drug, household, or other uses.
3.4 For your safety and health, please wear a lab coat and disposable gloves to operate.

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

270.33

C₂₀H₁₄O

5471-63-6

Solid

Light yellow to green yellow

523

495

C1(C2=CC=CC=C2)=C3C=CC=CC3=C(C4=CC=CC=C4)O1

Room temperature in continental US; may vary elsewhere.

4°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Bai X, et al. HKOH-1: A Highly Sensitive and Selective Fluorescent Probe for Detecting Endogenous Hydroxyl Radicals in Living Cells. Angew Chem Int Ed Engl. 2017 Oct 9;56(42):12873-12877.  [Content Brief]

  [2]. Weili Fan, et al. Calcium carbonate-methylene blue nanohybrids for photodynamic therapy and ultrasound imaging. Sci China Life Sci. 2018 Apr;61(4):483-491.  [Content Brief]

  [3]. P. Carloni, et al. On the use of 1,3-diphenylisobenzofuran (DPBF). Reactions with carbon and oxygen centered radicals in model and natural systems. Research on Chemical Intermediates volume 19, pages395-405(1993).