Real-time monitoring of redox changes in cells with a microplate reader
Can redox sensitive probes be used to monitor redox enzyme activity? To address this question, Prof. Morgan and his team developed a series of roGFP2 fusion constructs. roGFP (redox-sensitive GFP) is a modified version of GFP whose fluorescence properties are dependent on its redox state. By combining roGFP2 with glutaredoxin or glutathione the team was able to induce the oxidation of the signal molecule. The change of roGFP fluorescence can be monitored in semi-high-throughput on a CLARIOstar Plus microplate reader which opens up interesting new possibilities for the investigation of the cellular redox environment and reactive oxygen species generation and regulation.