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Related Experiment Videos

Abscisic Acid ELISA: Organic Acid Interference.

H Belefant1, F Fong

  • 1Department of Soil and Crop Sciences, Texas Agricultural Experiment Station, Texas A & M University System, College Station, Texas 77843.

Plant Physiology
|December 1, 1989
PubMed
Summary

False-positives in enzyme-linked immunosorbent assays (ELISAs) for abscisic acid (ABA) can occur due to organic acids and buffers. Careful selection of assay components is crucial for accurate ABA quantification.

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Area of Science:

  • Biochemistry
  • Plant Physiology
  • Analytical Chemistry

Background:

  • Enzyme-linked immunosorbent assays (ELISAs) are widely used for quantifying plant hormones like abscisic acid (ABA).
  • Accurate ABA measurement is critical for understanding plant stress responses and development.
  • Potential interferences in ELISA kits can compromise data reliability.

Purpose of the Study:

  • To investigate the causes of false-positive results in a commercial monoclonal antibody-based ELISA kit for abscisic acid (ABA).
  • To identify specific interfering substances among tricarboxylic acid cycle intermediates and common organic buffers.
  • To develop a method for removing interfering contaminants and ABA from commercial materials.

Main Methods:

  • ELISA assays were performed using a commercial monoclonal antibody kit for ABA.

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  • Standard curves of ABA were analyzed in the presence of various organic acids (tricarboxylic acid cycle intermediates) and organic buffers (Tris, Tricine, Hepes).
  • Interference was assessed by observing deviations in the slopes of standard curves.
  • A commercial sample of mannitol was tested for the presence of ABA.

    • Main Results:

    • A commercial ABA ELISA kit exhibited significant false-positive signals when tested with tricarboxylic acid cycle intermediates.
    • Organic acids and contaminants interfered with ABA quantification, altering standard curve slopes.
    • Alpha-ketoglutarate interference was attributed to a contaminant.
    • Among tested organic buffers, only Hepes induced false-positive ABA signals.
    • Commercial mannitol was found to contain ABA, necessitating a purification procedure.

    Conclusions:

    • The choice of buffers and solutions is critical for accurate ELISA-based ABA quantification.
    • Certain organic acids and buffers can cause significant interference, leading to false-positive ABA results.
    • A simple procedure for removing ABA from commercial mannitol is presented, improving its suitability for assays.