Immunoaffinity Purification of Indole-3-acetamide Using Monoclonal Antibodies
- 1 July 1987
- journal article
- research article
- Published by Oxford University Press (OUP) in Plant and Cell Physiology
- Vol. 28 (5) , 937-945
- https://doi.org/10.1093/oxfordjournals.pcp.a077376
Abstract
A single-step immunoaffinity purification procedure using monoclonal antibodies was developed to isolate indole-3-acetamide from plant extracts. Antibodies from a selected clone, raised against IAA-Cl′-BSA, with pronounced ability to recognize indole-3-acetamide (IAM) were used to prepare an immunoaffinity absorbent. Antibodies purified by thiophilic interaction chromatography were immobilized on divinylsulfoneactivated agarose. This column shows a very high selectivity towards IAM compared to IAA. This single step of immunoaffinity purification gave plant extracts of sufficient purity for direct quantification by on-line spectrofluorimetry after an analytical ionsuppression-HPLC run. Successive approximation by a second analytical ion-pairing-HPLC run confirmed the validity of this analytical technique.Keywords
This publication has 14 references indexed in Scilit:
- T-DNA-encoded auxin formation in crown-gall cellsPlanta, 1985
- Crown gall oncogenesis: evidence that a T-DNA gene from the Agrobacterium Ti plasmid pTiA6 encodes an enzyme that catalyzes synthesis of indoleacetic acid.Proceedings of the National Academy of Sciences, 1984
- Genetic analysis of the individual T-DNA genes of Agrobacterium tumefaciens; further evidence that two genes are involved in indole-3-acetic acid synthesisMolecular Genetics and Genomics, 1984
- The T‐region of Ti plasmids codes for an enzyme synthesizing indole‐3‐acetic acidEuropean Journal of Biochemistry, 1984
- Cloning characterization of iaaM, a virulence determinant of Pseudomonas savastanoi.Journal of Bacteriology, 1982
- Radioimmunoassay for pmol-quantities of indole-3-acetic acid for use with highly stable [125I]- and [3H]IAA derivatives as radiotracersPlanta, 1981
- A specific radioimmunoassay for nanogram quantities of the auxin, indole-3-acetic acidPlanta, 1977
- MICROBIAL SYNTHESIS AND DEGRADATION OF INDOLE-3-ACETIC ACID .I. CONVERSION OF L-TRYPTOPHAN TO INDOLE-3-ACETAMIDE BY AN ENZYME SYSTEM FROM PSEUDOMONAS SAVASTANOI1966
- Organic Growth Factor Requirements of Tobacco Tissue CulturesPhysiologia Plantarum, 1965
- Indoleacetamide as an Intermediate in the Synthesis of Indoleacetic Acid in Pseudomonas savastanoiScience, 1963