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| Dr. Rafal Nazarewicz |
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Plate reader |
FRIMCORE is equipped with new state-of-the-art high sensitivity fluorescence plate reader Synergy H1 (BioTeck) with monochrome-based fluorescent spectrometer and temperature controlled sampler. We perform analysis of hydrogen peroxide using Amplex Red assay and superoxide measurements (DHE or MitoSOX) in 24 to 96 well based samples. Our new methods include analysis of nitric oxide production (DAF2-DA) and measurements of peroxynitrite (CBA) in cellular and tissue samples. The main advantage of these methods is high throughput while superoxide measurements by HPLC and nitric oxide detection by EPR provide more specific and quantitative analysis. In cases of small cell number or high sample number, however, fluorescent monochromatic based plate reader is a good alternative.
The following plate-reader services are provided:
1) Measurement of cellular superoxide by DHE;
2) Measurement of mitochondrial superoxide by MitoSOX;
3) Measurement of cellular nitric oxide production by DAF2-DA;
4) Measurement of measurements of peroxynitrite by CBA.
We recommend scheduling a meeting prior to plate reader based analysis in order to determine the best protocol for sample preparation. This service may require training for sample preparation; however, all samples will be processed by FRIMCORE staff. To schedule a meeting or consultation please contact Dr. Sergey Dikalov or Rafal Nazarewicz:
- Dikalov SI, Harrison DG. Methods for Detection of Mitochondrial and Cellular Reactive Oxygen Species. Antioxid Redox Signal. 2012.
- Zielonka J, Sikora A, Hardy M, Joseph J, Dranka BP, Kalyanaraman B.Boronate probes as diagnostic tools for real time monitoring of peroxynitrite and hydroperoxides. Chem Res Toxicol. 2012;25(9):1793-9.
- Zielonka J, Zielonka M, Sikora A, Adamus J, Joseph J, Hardy M, Ouari O,Dranka BP, Kalyanaraman B. Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem. 2012; 287(5):2984-95.
- Nazarewicz RR, Bikineyeva A, Dikalov SI. Rapid and specific measurements of superoxide using fluorescence spectroscopy. JBiomolecular Screening 2012
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