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

Abstract

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.

Authors:
; ; ; ;
Publication Date:
Research Org.:
Queen's Univ. of Belfast (England)
OSTI Identifier:
6957019
Alternate Identifier(s):
OSTI ID: 6957019
Resource Type:
Journal Article
Resource Relation:
Journal Name: Plant Physiol.; (United States); Journal Volume: 85:1
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ABSCISIC ACID; ENZYME IMMUNOASSAY; GAS CHROMATOGRAPHY; LABELLED COMPOUNDS; LEAVES; MASS SPECTROSCOPY; MONOCLONAL ANTIBODIES; PISUM; QUANTITATIVE CHEMICAL ANALYSIS; SENSITIVITY; TRITIUM COMPOUNDS; VALIDATION; ANTIBODIES; AUXINS; BACTERIA; CARBOXYLIC ACIDS; CHEMICAL ANALYSIS; CHROMATOGRAPHY; IMMUNOASSAY; LEGUMINOSAE; MICROORGANISMS; MONOCARBOXYLIC ACIDS; ORGANIC ACIDS; ORGANIC COMPOUNDS; PLANT GROWTH REGULATORS; PLANTS; RHIZOBIUM; SEPARATION PROCESSES; SPECTROSCOPY; TESTING 550200* -- Biochemistry

Citation Formats

Ross, G.S., Elder, P.A., McWha, J.A., Pearce, D., and Pharis, R.P.. Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum]. United States: N. p., 1987. Web.
Ross, G.S., Elder, P.A., McWha, J.A., Pearce, D., & Pharis, R.P.. Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum]. United States.
Ross, G.S., Elder, P.A., McWha, J.A., Pearce, D., and Pharis, R.P.. Tue . "Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum]". United States.
@article{osti_6957019,
title = {Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum]},
author = {Ross, G.S. and Elder, P.A. and McWha, J.A. and Pearce, D. and Pharis, R.P.},
abstractNote = {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.},
doi = {},
journal = {Plant Physiol.; (United States)},
number = ,
volume = 85:1,
place = {United States},
year = {Tue Sep 01 00:00:00 EDT 1987},
month = {Tue Sep 01 00:00:00 EDT 1987}
}