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Method Article
A high throughput spectrophotometric method for singlet oxygen quenching
Suaib Luqman
Suaib Luqman
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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
- Ashleigh Carver, Editorial Assistant, Nature Protocols
Keywords
Singlet oxygen, Hypochlorous acid, Hydrogen peroxide, RNO, L-Histidine
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Troubleshooting Limitations
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Tables 1-3 Tables
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Supplementary Document 1 Version 1 of Protocol Edited 29/07/2016 - Ashleigh Carver, Editorial Assistant, Nature Protocols
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Associated Publications
Quenching of singlet oxygen by natural and synthetic antioxidants and assessment of electronic UV/Visible absorption spectra for alleviating or enhancing the efficacy of photodynamic therapy
by Kaneez Fatima, Nusrat Masood, and Suaib Luqman
Eclipta yellow vein virus enhances chlorophyll destruction, singlet oxygen production and alters endogenous redox status in Andrographis paniculata
by Asifa Khan, Suaib Luqman, Nusrat Masood, +3
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This is a preprint. It has not completed peer review.
Method Article
A high throughput spectrophotometric method for singlet oxygen quenching
Suaib Luqman
Suaib Luqman
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 26 Jul, 2016
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biochemistry
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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
- Ashleigh Carver, Editorial Assistant, Nature Protocols
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Associated Publications
Quenching of singlet oxygen by natural and synthetic antioxidants and assessment of electronic UV/Visible absorption spectra for alleviating or enhancing the efficacy of photodynamic therapy
by Kaneez Fatima, Nusrat Masood, and Suaib Luqman
Eclipta yellow vein virus enhances chlorophyll destruction, singlet oxygen production and alters endogenous redox status in Andrographis paniculata
by Asifa Khan, Suaib Luqman, Nusrat Masood, +3