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Assay of the polyamine spermine by a monoclonal antibody-based ELISA
I Garthwaite1, A D Stead, C C Rider
1Department of Biochemistry, University of London, Egham, Surrey, UK.
Abstract:
Spermine-specific monoclonal antibodies were prepared by immunising mice with protein-spermine conjugates. A resulting monoclonal antibody, IAG-1, exhibited both high affinity for spermine (binding constant 5.5 x 10(7) M-1) and high specificity, cross-reacting only weakly with spermidine. Using this antibody a competitive ELISA was developed with a detection limit of 1 pmol. The assay has been used to quantify spermine content of plant tissues, without derivatization, and producing within 6 h of collection, values for the spermine content which are similar to published data obtained by HPLC.
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