Characterization of methylsulfinylalkyl glucosinolate specific polyclonal antibodies
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Antibodies towards small molecules, like plant specialized metabolites, are valuable tools for developing quantitative and qualitative analytical techniques. Glucosinolates are the specialized metabolites characteristic of the Brassicales order. Here we describe the characterization of polyclonal rabbit antibodies raised against the 4-methylsulfinylbutyl glucosinolate, glucoraphanin that is one of the major glucosinolates in the model plant Arabidopsis thaliana (hereafter Arabidopsis). Analysis of the cross-reactivity of the antibodies against a number of glucosinolates demonstrated that it was highly selective for methionine-derived aliphatic glucosinolates with a methyl-sulfinyl group in the side chain. Use of crude plant extracts from Arabidopsis mutants with different glucosinolate profiles showed that the antibodies recognized aliphatic glucosinolates in a plant extract and did not cross-react with other metabolites. These methylsulfinylalkyl glucosinolate specific antibodies have prospective use in multiple applications such as ELISA, co-immunoprecipitation and immunolocalization of glucosinolates.
KeywordsAntibody characterization Small molecules Methylsulfinylalkyl glucosinolate
This study is financially supported by the Danish National Research Foundation (grant DNRF99). We wish to thank Dr. Frederikke Gro Malinovsky from the DynaMo Center, University of Copenhagen for her helpful ideas and discussions in the preparation of this manuscript.
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