A high throughput spectrophotometric method for singlet oxygen quenching
An easy spectrophotometric method for evaluating quenchers of singlet oxygen is described which can be used for high throughput assessment of large number of samples in a 96-well plate format. Singlet oxygen is a higher energy state molecular oxygen species produced in various intracellular reactions, extracellular reactions by the aid of photo sensitizer, light source and/or by chemical means. In this protocol, we have developed a simple and accurate procedure for generating singlet oxygen with no side reactions at physiological pH. The second aspect of this assay is detection by p-nitroso dimethylaniline (RNO), which is an already established suitable trap with imidazole and/or its derivatives for singlet oxygen. We have synchronized the assay system by performing various combinations of reagent used and also by plotting their spectra, pH and time-dependence curve monitored spectrophotometrically at 440nm
COMMENTS
Protocol edited 29/07/2016
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Supplementary Document 1 Version 1 of Protocol Edited 29/07/2016 - Ashleigh Carver, Editorial Assistant, Nature Protocols
Posted 26 Jul, 2016
A high throughput spectrophotometric method for singlet oxygen quenching
Posted 26 Jul, 2016
An easy spectrophotometric method for evaluating quenchers of singlet oxygen is described which can be used for high throughput assessment of large number of samples in a 96-well plate format. Singlet oxygen is a higher energy state molecular oxygen species produced in various intracellular reactions, extracellular reactions by the aid of photo sensitizer, light source and/or by chemical means. In this protocol, we have developed a simple and accurate procedure for generating singlet oxygen with no side reactions at physiological pH. The second aspect of this assay is detection by p-nitroso dimethylaniline (RNO), which is an already established suitable trap with imidazole and/or its derivatives for singlet oxygen. We have synchronized the assay system by performing various combinations of reagent used and also by plotting their spectra, pH and time-dependence curve monitored spectrophotometrically at 440nm
COMMENTS
Protocol edited 29/07/2016
Figure 1
Figure 2
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Figure 7
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