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Bio Protoc. 2012;2(18). pii: e263. Epub 2012 Sep 20.

Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves.

Author information

1
Department of Biological Sciences, Royal Holloway University of London, Egham, UK; Department of Plant Pathology, University of California, Davis, CA, USA.
2
Department of Biological Sciences, Royal Holloway University of London, Egham, UK.

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.

PMID:
27390754
PMCID:
PMC4932902

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