Abstract
The development of a sensitive and specific enzyme immunoassay for GA3 is reported. This method was based on the use of peroxidase labelled GA3 and immobilized antibodies. In order to obtain a rapid immunoassay, several steps of purification were analyzed to show their necessity. Barley seed extracts were assayed at different steps of purification to exhibit the effect of extract components on the assay. It was demonstrated that HPLC had to be performed when a selective quantitation of GA3 was required. This assay allowed GA3 to be measured with reproducibility as its unmethylated form and the quantitation of GA3 in barley seeds with this enzyme immunoassay was correlated to a GC-MS method.
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Abbreviations
- GA3 :
-
gibberellin A3
- EIA:
-
enzyme immunoassay
- DMF:
-
dimethylformamide
- TEA:
-
tri(n)ethylamine
- BSA:
-
bovine serum albumin
- OVA:
-
ovalbumine
- ECF:
-
ethylchloroformate
- PB:
-
phosphate buffer
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Bounaix, C., Doumas, P. Development of a one-step enzyme immunoassay for the quantitation of gibberellin A3 in barley seeds. Plant Growth Regul 17, 7–13 (1995). https://doi.org/10.1007/BF00024488
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DOI: https://doi.org/10.1007/BF00024488