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Does anyone have a good protocol/kit for measuring ROS levels in...
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+ Molecular plant physiology
+ Plant biotechnology
+ Reactive oxygen species
+ Plant molecular biology
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Milton Steinberg
Does anyone have a good protocol/kit for measuring ROS levels in...
This is a very similar question to other post, I just contributed to. I think it would be good if you could narrow down your question. Which type of ROS? In which organelle you think it is formed? There are many sites where ROS are generated and accumulated. Have in mind that because you are working with plants, photoinduced ROS will be always in the background. Because ROS are very reactive, their lifetime are too short. So most in methods you will find are indirect and could be biased. DAB is good for peroxide, but most of the time peroxide is a side product and it is a quite messy technique. Fluorescent methods are very common but they have problems because in any fluorescence methods you may have quenchers that would cause artifacts. If you are isolating organelles you could try EPR using spin traps (such as EMPO and TEMPO). Other option is singlet phosphorescence in the NIR to detect singlet oxygen (by the way this a direct method, you really measure singlet oxygen). Just keep in mind that you should measure them in darkness, because any stressed induced in the tissue will enhance the photo-induced ROS and then you will measure something else. (non-proofread)
This is a very similar question to other post, I just contributed to. I think it would be good if you could narrow down your question. Which type of ROS? In which organelle you think it is formed? There are many sites where ROS are generated and accumulated. Have in mind that because you are working with plants, photoinduced ROS will be always in the background. Because ROS are very reactive, their lifetime are too short. So most in methods you will find are indirect and could be biased. DAB is good for peroxide, but most of the time peroxide is a side product and it is a quite messy technique. Fluorescent methods are very common but they have problems because in any fluorescence methods you may have quenchers that would cause artifacts. If you are isolating organelles you could try EPR using spin traps (such as EMPO and TEMPO). Other option is singlet phosphorescence in the NIR to detect singlet oxygen (by the way this a direct method, you really measure singlet oxygen). Just keep in mind that you should measure them in darkness, because any stressed induced in the tissue will enhance the photo-induced ROS and then you will measure something else. (non-proofread)
This protocol might be of help: Juszczak I., Baier M. (2014) Quantification of Superoxide and Hydrogen Peroxide in Leaves. In: Hincha D., Zuther E. (eds) Plant Cold Acclimation. Methods in Molecular Biology (Methods and Protocols), vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_16
This protocol might be of help: Juszczak I., Baier M. (2014) Quantification of Superoxide and Hydrogen Peroxide in Leaves. In: Hincha D., Zuther E. (eds) Plant Cold Acclimation. Methods in Molecular Biology (Methods and Protocols), vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_16
As lipid peroxidation is considered as an indicator of oxidative stress, malondialdehyde (MDA) measurement (they are produced from lipid peroxides) can give you an estimate for oxidative stress and ROS.
As lipid peroxidation is considered as an indicator of oxidative stress, malondialdehyde (MDA) measurement (they are produced from lipid peroxides) can give you an estimate for oxidative stress and ROS.
This is a very similar question to other post, I just contributed to. I think it would be good if you could narrow down your question. Which type of ROS? In which organelle you think it is formed? There are many sites where ROS are generated and accumulated. Have in mind that because you are working with plants, photoinduced ROS will be always in the background.
Because ROS are very reactive, their lifetime are too short. So most in methods you will find are indirect and could be biased. DAB is good for peroxide, but most of the time peroxide is a side product and it is a quite messy technique. Fluorescent methods are very common but they have problems because in any fluorescence methods you may have quenchers that would cause artifacts. If you are isolating organelles you could try EPR using spin traps (such as EMPO and TEMPO). Other option is singlet phosphorescence in the NIR to detect singlet oxygen (by the way this a direct method, you really measure singlet oxygen). Just keep in mind that you should measure them in darkness, because any stressed induced in the tissue will enhance the photo-induced ROS and then you will measure something else.
(non-proofread)
This is a very similar question to other post, I just contributed to. I think it would be good if you could narrow down your question. Which type of ROS? In which organelle you think it is formed? There are many sites where ROS are generated and accumulated. Have in mind that because you are working with plants, photoinduced ROS will be always in the background.
Because ROS are very reactive, their lifetime are too short. So most in methods you will find are indirect and could be biased. DAB is good for peroxide, but most of the time peroxide is a side product and it is a quite messy technique. Fluorescent methods are very common but they have problems because in any fluorescence methods you may have quenchers that would cause artifacts. If you are isolating organelles you could try EPR using spin traps (such as EMPO and TEMPO). Other option is singlet phosphorescence in the NIR to detect singlet oxygen (by the way this a direct method, you really measure singlet oxygen). Just keep in mind that you should measure them in darkness, because any stressed induced in the tissue will enhance the photo-induced ROS and then you will measure something else.
(non-proofread)
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We usually use the protocols written in:
Article Comparative analysis of overexpressed Fragaria vesca S-adeno...
We usually use the protocols written in:
Article Comparative analysis of overexpressed Fragaria vesca S-adeno...
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many thanks dear zaynab but i mean accurate measurement of ROS like HO2O or another ROS molecules ...
many thanks dear zaynab but i mean accurate measurement of ROS like HO2O or another ROS molecules ...
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Check my article for Catalase and Peroxidase enzyme activities.Article Impact of water-soluble 2D CdSe/CdS nanoplatelets on seed ge...
Check my article for Catalase and Peroxidase enzyme activities.Article Impact of water-soluble 2D CdSe/CdS nanoplatelets on seed ge...
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Check out the protocol described in "Rapid bioassay to measure early reactive oxygen species production in Arabidopsis leave tissue in response to living Pseudomonas syringae" by Smith & Heese (2014):
http://plantmethods.biomedcentral.com/articles/10.1186/1746-4811-10-6
Also consider detecting ROS by DAB (3,3-diaminobenzidine) or NBT (nitroblue tetrazolium) staining (See links below):
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1475455/
http://www.bio-protocol.org/e1108
Check out the protocol described in "Rapid bioassay to measure early reactive oxygen species production in Arabidopsis leave tissue in response to living Pseudomonas syringae" by Smith & Heese (2014):
http://plantmethods.biomedcentral.com/articles/10.1186/1746-4811-10-6
Also consider detecting ROS by DAB (3,3-diaminobenzidine) or NBT (nitroblue tetrazolium) staining (See links below):
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1475455/
http://www.bio-protocol.org/e1108
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This protocol might be of help:
Juszczak I., Baier M. (2014) Quantification of Superoxide and Hydrogen Peroxide in Leaves. In: Hincha D., Zuther E. (eds) Plant Cold Acclimation. Methods in Molecular Biology (Methods and Protocols), vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_16
This protocol might be of help:
Juszczak I., Baier M. (2014) Quantification of Superoxide and Hydrogen Peroxide in Leaves. In: Hincha D., Zuther E. (eds) Plant Cold Acclimation. Methods in Molecular Biology (Methods and Protocols), vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_16
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Go to Plantstress.com. Click on 'Methods'. Check the list for "Oxidative".
Go to Plantstress.com. Click on 'Methods'. Check the list for "Oxidative".
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Go to Plantstress.com. Click on 'Methods'. Check the list for "Oxidative".
Go to Plantstress.com. Click on 'Methods'. Check the list for "Oxidative".
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As lipid peroxidation is considered as an indicator of oxidative stress, malondialdehyde (MDA) measurement (they are produced from lipid peroxides) can give you an estimate for oxidative stress and ROS.
As lipid peroxidation is considered as an indicator of oxidative stress, malondialdehyde (MDA) measurement (they are produced from lipid peroxides) can give you an estimate for oxidative stress and ROS.
More
VOTE