Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum]

Journal Article · Tue Sep 01 04:00:00 EDT 1987 · Plant Physiol.; (United States)

OSTI ID:6957019

Ross, G S; Elder, P A; McWha, J A; Pearce, D; Pharis, R P

AN INDIRECT METHOD OF ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA) IS REPORTED FOR ABSCISIC ACID (ABA), UTILIZING A THYROGLOBULIN-ABA CONJUGATE FOR COATING WELLS. THE ASSAY CAN USE COMMERCIALLY AVAILABLE MONOCLONAL ANTIBODIES, IS SENSITIVE TO AS LITTLE AS 20 PICOGRAMS ABA PER WELL, AND IS MUCH MORE CONSERVATIVE OF ANTIBODY THAN DIRECT METHODS. THE MOST DILUTE ABA STANDARDS DID NOT RETAIN THEIR ANTIGENICITY DURING STORAGE, SO ABA STANDARD SETS WERE DILUTED IMMEDIATELY PRIOR TO USE. THE INDIRECT ELISA WAS USED SUCCESSFULLY TO ESTIMATE ABA CONCENTRATIONS IN DEVELOPING COTYLEDONS OF PISUM SATIVUM L., AFTER ONLY LITTLE PRELIMINARY PURIFICATION. IT WAS VALIDATED FOR THIS TISSUE THROUGH THE USE OF GAS CHROMATOGRAPHY-ELECTRON CAPTURE DETECTION (GC-EC), AND CAPILLARY GC-SELECTED ION MONITORING (GC-MS-SIM) USING LABELLED ABA AS AN INTERNAL STANDARD. FULL SPECTRUM GC-MASS SPECTROMETRY WAS ALSO USED TO VERIFY THAT ABA WAS PRESENT IN A SAMPLE ASSAYED QUANTITATIVELY BY BOTH ELISA AND GC-MS-SIM.

• Research Organization: Queen's Univ. of Belfast (England)
• OSTI ID: 6957019
• Journal Information: Plant Physiol.; (United States), Journal Name: Plant Physiol.; (United States) Vol. 85:1; ISSN PLPHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ABSCISIC ACID
ANTIBODIES
AUXINS
BACTERIA
CARBOXYLIC ACIDS
CHEMICAL ANALYSIS
CHROMATOGRAPHY
ENZYME IMMUNOASSAY
GAS CHROMATOGRAPHY
IMMUNOASSAY
LABELLED COMPOUNDS
LEAVES
LEGUMINOSAE
MASS SPECTROSCOPY
MICROORGANISMS
MONOCARBOXYLIC ACIDS
MONOCLONAL ANTIBODIES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PISUM
PLANT GROWTH REGULATORS
PLANTS
QUANTITATIVE CHEMICAL ANALYSIS
RHIZOBIUM
SENSITIVITY
SEPARATION PROCESSES
SPECTROSCOPY
TESTING
TRITIUM COMPOUNDS
VALIDATION