Hi, ArsalanThanks for your protocol.But I have two questions.1....

WB
Wenlu Bi
Jul 23, 2015
protocol Protocol: Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves
Hi, Arsalan
Thanks for your protocol.
But I have two questions.
1. If i want to make a 1 L DAB solution, how much water could be used? if i could added water to the powder until the volume is 1 liter?
2. About the bleaching solution. what do you mean by Ethanol: Acetic
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Wenlu Bi Answered Jul 23, 2015

NWAFU

Thanks a lot for your answer. It is very helpful. I will try again as you recommend !

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Arsalan Daudi Author Answered Jul 23, 2015

University of California

Hi Wenlu,
Thanks for your message.

To make 1 L DAB solution, I would recommend adding 900 ml sterile distilled water to 1 g DAB, then add proportional Tween-20 and top up with 200 mM Na2HPO4. Also, try to use acidified DAB instead of just DAB, then you don't need to do the acidification step in my protocol.

Yes, Ethanol: Acetic Acid: Glycerol=3:1:1 means 3 portion ethanol, 1 portion so on. Don't make it in water. Take 300 ml Ethanol, add 100 ml Acetic acid and 100 ml glycerol, and use this as the bleaching solution.

It would be great if you post your result here. Good luck!
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