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

Bruno Sotta | Zendy; Gilles Pilate | Zendy; Florence Pelèse | Zendy; I. Sabbagh | Zendy; Malgorzata Bonnet | Zendy; Régis Maldiney | Zendy

Open Access

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

Author(s) -

Bruno Sotta,

Gilles Pilate,

Florence Pelèse,

I. Sabbagh,

Malgorzata Bonnet,

Régis Maldiney

Publication year - 1987

Publication title -

plant physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.554

H-Index - 312

eISSN - 1532-2548

pISSN - 0032-0889

DOI - 10.1104/pp.84.3.571

Subject(s) - biotinylation , avidin , chromatography , chemistry , high performance liquid chromatography , immunoassay , conjugate , biotin , quantitative analysis (chemistry) , antibody , alkaline phosphatase , biochemistry , enzyme , biology , immunology , mathematical analysis , mathematics

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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