Nature Protocols Discussion Forum: topic
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hydrogen peroxide detection
Jyothilakshmi V
Wednesday, 02 May 2007 15:42 UTC
does anybody know how reliable is nbt staining for h2o2 detection in arabidopsis..
also to do fluorescent staining by DCF ..how to avoivd background
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Replies
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Hi, for measuring the peroxide with DCF you should use very low concentration of the dye.Also, you can use some recombinant fluorescent proteins to do so.See the paper published in Nature Methods(www.nature.com/nmeth/journal/v3/n4/abs/nmeth866.html)
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I believe NBT isn’t very sensitive and has a certain threshold when the cell becomes completely positive. At least, that’s what we see in blood-cells. So it’s a black/white screening and not always points out that there isn’t any H2O2 production if the cell is negative.
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