FBBBE 

FBBBE is used to detect the production of H₂O₂ by cells. FBBBE can be triggered by intracellular H₂O₂ and converted to fluorescein, resulting in an increase in intracellular fluorescence (Ex=4480 nm, Em=512 nm)^[1].

H₂O₂^[1]

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1].
1. The RAW 264.7 cells are seeded at 250,000 in glass bottom 35 mm dishes and post-adhering (5 h).
2. Some dishes are treated with 1 μg/mL of LPS for 24 h. Other dishes are treated the next day after plating with either 100 μM H₂O₂ for 1 h or PBS for 1 h.
3. Cells are washed twice with PBS and fixed in ice-cold 95% ethanol for 15 min.
3. Cells are twice rinsed with PBS and incubated with either 50 μM probe (FBBBE) for 1 h at room temperature.
4. Cells are washed twice with PBS and coverslips were mounted over cells using Aqua Mount.
5. Confocal images are obtained at 80× magnification on an Olympus FV1000 confocal laser scanning microscope.
6. Images were processed using ImageJ software.

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

764.47

C₄₆H₄₆B₂O₉

1522117-83-4

Solid

Off-white to light yellow

512

480

O=C1OC2(C3=C1C=CC=C3)C4=CC=C(C=C4OC5=CC(OCC6=CC=C(B7OC(C)(C(C)(C)O7)C)C=C6)=CC=C52)OCC8=CC=C(B9OC(C)(C(C)(C)O9)C)C=C8

Room temperature in continental US; may vary elsewhere.

-20°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]. Daniel, K.B., et al. Readily accessible fluorescent probes for sensitive biological imaging of hydrogen peroxide. Chembiochem 14(5), 593-598 (2013).