ROS Facility
Reactive oxygen species (ROS) are free radical and non-free radical oxygen intermediates formed from the natural metabolism of oxygen that function to oxidize their cellular targets. ROS are generated by a variety of enzymes including NADPH oxidases (Nox), xanthine oxidase, mitochondrial enzymes, and dysfunctional nitric oxide synthases. Once thought to subserve a more narrow role in phagocyte antimicrobial action, the degradation of bioactive nitric oxide (NO), as well as cell damage as a result of 'oxidative stress', much evidence has emerged in recent years supporting a well-regulated signaling role for ROS in a variety of cell processes. These include among others cell proliferation, hypertrophy, migration, wound healing, neuronal signaling, vasomotor tone, and thyroid hormone metabolism. Thus there is broad interest in reliable and sensitive methods to measure the contribution of ROS to these and other processes at the cellular level.
The ROS Facility has been set up to aid investigators in the detection of ROS in multiple model systems, and is experienced in the following assays:
Plate-based Assays* of whole cells in culture using spectrophotometric, chemiluminescence and fluorescence techniques for detection and quantification of intracellular and extracellular:
superoxide anion
hydrogen peroxide
EPR (Electron Paramagnetic Resonance) Spectrometry
For identification and quantification of various ROS and NO. Using both spin traps/probes and direct radical detection methods, ROS and NO formation are determined in cellular systems and in ex vivo/in vivo samples allowing for enhanced specificity and verification of alternative methods.
- Intracellular and extracellular ROS detection
- Cellular NO formation
- Tissue ROS and NO formation
- Tissue oxygen concentration measurements
- Hydrogen peroxide detection
*Plate- and tube-based assays are currently being developed to measure ROS at the subcellular and tissue level and are expected to be available soon.
The staff of the ROS Facility will provide consultation to researchers in the design and interpretation of experiments involving ROS detection.
For more information, please contact Patrick Pagano, Ph.D.
