Plant Physiol.
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Plant Physiology 84:571-573 (1987)
© 1987 American Society of Plant Biologists

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Development and Growth Regulation

An Avidin-Biotin Solid Phase ELISA for Femtomole Isopentenyladenine and Isopentenyladenosine Measurements in HPLC Purified Plant Extracts

Bruno Sotta, Gilles Pilate, Florence Pelese, Issam Sabbagh, Malgorzata Bonnet and Regis Maldiney

Laboratoire de Physiologie Végétale, T 53, 5ème étage, Université P et M Curie, 4 place Jussieu, 75252 Paris Cedex 05, France, Institut Pasteur Lyon, Domaine du Poirier, Lentilly, 69210 L'Arbresles, France

A solid phase enzyme immunoassay was developed for isopentenyladenine (iP) and isopentenyladenosine (iPA) quantitation in HPLC purified plant extracts. It was performed on antigen-coated microtitration plates on which bound antibodies were indirectly labeled by the means of a biotinylated goat anti-rabbit antibody and an avidin-alkaline phosphatase conjugate. Less than 3 femtomoles of iP or iPA were easily detected and the measuring range extended from 3 femtomole to 1 picomole. The reproducibility has been tested and intra- and interassay variations did not exceed 5.0%. The specificity of iPA antibodies was good, as determined by cross-reactivity measurements with other adenylic compounds. The specificity of the measurements for iP and iPA was demonstrated by analysis of the immunoreactivity of fractions obtained by HPLC separation of a methanolic tobacco leaf extract.






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ASPB Publications PLANT PHYSIOLOGY THE PLANT CELL
Copyright © 1987 by the American Society of Plant Biologists