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].

Research Areas >> Cancer

Signaling Pathways >> Others >> Others

Research Areas >> Inflammation/Immunology

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.

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