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Chemiluminescence measurement of the generation of reactive oxygen species

Bernd Hipler (1), Uta-Christina Hipler (2) (1) BMG LABTECH, (2) FSU Jena 12/2014

During the unspecific immune response, oxygen demand is strongly increased as oxygen is converted into superoxide anions, hydrogen peroxide, monomolecular oxygen and hydroxyl c radicals. These molecules are released in a respiratory burst to harm bacteria, parasites and tumor cells. These extracellular highly reactive oxygen species (ROS) cause many biological effects such as destruction.

 

Lucigenin reacts with superoxide under emission of light. In this application note we show the measurement of ROS generation in the yeast Candida albicans using lucigenin. In C. albicans, mitochondria are capable of generating extracellular released ROS. The effect of terbinafine, a considered free radical interceptor, on ROS formation is investigated.

 

The chemiluminescent signal was measured on a BMG LABTECH microplate reader and was sensitive enough to detect low ROS concentrations of unstimulated C. albicans. Furthermore, it was shown that terbinafine not only has radical scavenging properties, but also interacts with the pathway that generates reactive oxygen species.

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