
Background: This ELISA assay utilizes the principle of competitive binding to measure the concentration of hormone in plant extracts. The ABA (abscisic acid) hormone has been pre-coated on the surface of the reaction wells. The plant extract sample, containing an unknown amount of hormone, or standards are mixed in the reaction well with a known amount of antibody to ABA. During incubation the competitive inhibition reaction occurs between pre-coated ABA and hormone in the samples with the antibody specific to ABA.
Unbound hormone and plant extract are washed out of the reaction wells. Addition of HRP-conjugated goat anti-rabbit IgG antibody will visualize binding between anti-ABA antibodies and ABA hormone. Substrate solution is added to the wells and the color develops in opposite to the amount of ABA in the sample or standards. Reaction is stopped and the intesity of the color is measured at 450 nm.
Reaction wells: 5x96 wells
Assay development time: 1-2 hours
Sensitivity: 0.04 μg/ml
Detection ragne: 0.156 μg/ml-10 μg/ml
Plant extract volume: 50-100 µl
Detection wavelength: 450 nm
Intra-assay precision (within an assay): CV%<10%
Intra-assay precision (between assays): CV%<20%
Storage: 2-8°C
Reference: Cassani et al. (2024). Drought- and Salt-Tolerant Populations of the Xero-Halophyte Mediterranean Shrub Atriplex halimus L. Exhibit Contrasting Proline and Glycinebetaine Metabolism. J Plant Growth Regul (2024). https://doi.org/10.1007/s00344-024-11558-7.
Compound | Cross-reactivity (%)
Gibberelin: < 0.01 %
Indoleacetic acid: < 0.01 %
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