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A specific radioimmunoassay for nanogram quantities of the auxin, indole-3-acetic acid

✍ Scribed by William Pengelly; Frederick Meins

Publisher
Springer-Verlag
Year
1977
Tongue
English
Weight
691 KB
Volume
136
Category
Article
ISSN
0032-0935

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✦ Synopsis

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, a-and fi-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.

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