Hui Wang1, Jean Osterman1, Wenhui Zhou1, Poncho Meisenheimer1, John Shultz2, Sarah Duellman2, Jolanta Vidugiriene2, Gediminas Vidugiris2 and James Cali2
1 Promega Biosciences, Inc. 277 Granada Drive, San Luis Obispo, CA 93401
2 Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711
Consideration of the chemistry and biology of H2O2in cell culture:
• H2O2 has a relatively long half-life in solution and can diffuse in/out of the cell.
• Some compounds are easily oxidized in solution by air to produce H2O2 through a cell-independent mechanism.
• Several enzymes in the cell (e.g., monoamine oxidases (MAOs), D-amino acid oxidase, demethylases, etc.)directly produce H2O2 as a byproduct of their enzymatic reaction.
• Compounds, such as menadione and rotenone, that interfere with the electron transport chain of the mitochondria can induce H2O2 generation through the action of superoxide dismutase.
• Considerations important in determining a detection method for H2O2
• Ease of use
• Minimal false positive hits in the presence of library test compounds
• Adequate detection sensitivity
• Appropriate pH, temperature and buffer