Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves
拟南芥叶中过氧化氢的检测（DAB染色法）
发布: 2012年09月20日第2卷第18期 DOI: 10.21769/BioProtoc.263 浏览次数: 85834
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The authors used this protocol in:
Jan 2012
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Abstract
In this protocol, the in situ detection of hydrogen peroxide (one of several reactive oxygen species) is described in mature Arabidopsis rosette leaves by staining with 3,3'-diaminobenzidine (DAB) using an adaptation of previous methods (Thordal-Christensen et al., 1997; Bindschedler et al., 2006; Daudi et al., 2012). DAB is oxidized by hydrogen peroxide in the presence of some haem-containing proteins, such as peroxidases, to generate a dark brown precipitate. This precipitate is exploited as a stain to detect the presence and distribution of hydrogen peroxide in plant cells. The protocol can be modified slightly to detect hydrogen peroxide in different types of plant tissue.
Keywords: Oxidative Burst (氧化爆裂)
Reactive Oxygen Species (活性氧)
Hydrogen Peroxide (过氧化氢)
Disease Resistance (抗病性)
Hypersensitive Response (过敏反应)
Materials and Reagents
- Arabidopsis plants
- DAB non-acidified powder (Sigma-Aldrich, catalog number: D8001 )
- Tween 20 viscous liquid molecular biology grade (Sigma-Aldrich, catalog number: P9416 )
- Sodium phosphate (Na2HPO4) electrophoresis grade (Sigma-Aldrich, catalog number: S3397 )
- Aluminum foil
- HCl
- Na2HPO4
- Ethanol
- Acetic acid
- Glycerol
- DAB staining solution (see Recipes)
- 200 mM Na2HPO4 (see Recipes)
- Bleaching solution (see Recipes)
Equipment
- Arabidopsis growth chamber
- Magnetic stirrer and stirring bar
- pH meter
- 1 ml needless syringes
- 12-well microtiter plate
- Dessicator
- Shaker
- Water bath
- 50 ml Falcon tube
Procedure
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文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Daudi, A. and O’Brien, J. A. (2012). Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves. Bio-protocol 2(18): e263. DOI: 10.21769/BioProtoc.263.
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分类
植物科学 > 植物免疫 > 病害生物测定
生物化学 > 其它化合物 > 活性氧
植物科学 > 植物生理学 > 组织分析
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