A specific radioimmunoassay for nanogram quantities of the auxin,indole-3-acetic acid |
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Authors: | William Pengelly Frederick Meins Jr |
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Institution: | (1) Department of Biology, Princeton University, 08540 Princeton, NJ, USA;(2) Department of Botany, University of Illinois, 61801 Urbana, IL, USA;(3) Present address: Department of Botany, University of Illinois, 61801 Urbana, IL, USA |
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Abstract: | We have developed a specific radioimmunoassay RIA] for indole-3-acetic acid (IAA) in the 0.2 ng to 12 ng range which, in principle, can be extended to other indole auxins as well. Methods are presented for obtaining suitable antibody, for the RIA procedure, and for measuring IAA in methanolic extracts of plant tissues. Antibody specific for IAA was obtained from rabbits immunized with IAA bound to bovine serum albumin by formaldehyde treatment. In assays with this antibody, 2,4-dichlorophenoxyacetic acid and indoles structurally related to IAA reacted from 300- to 3000-fold less than did IAA itself. However, -and -naphthaleneacetic acid reacted significantly and hence interfered with the assay. Extracts of tobacco (Nicotiana tabacum L.) tissue were immunoassayed after partial purification by buffer-ether partition. Crown-gall tumor tissue, which is auxin-autotrophic, and pith tissue depleted of auxin by the diffusion method contained, respectively, 26.7 ng and <0.5 ng extractable IAA per gram fresh weight.Abbreviations BSA
bovine serum albumin
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- -NAA
-naphthalenacetic acid
- PBS
phosphate-buffered saline
- RIA
radioimmunoassay |
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Keywords: | Auxin Immunoassay Nicotiana Radioimmunoassay |
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