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Preparation of iodine 125 labeled synthetic porcine secretin for radio immunoassay


, : Preparation of iodine 125 labeled synthetic porcine secretin for radio immunoassay. Scandinavian Journal of Clinical and Laboratory Investigation 36(7): 661-668

Synthetic porcine secretin was labeled by the conjugation-labeling method of Bolton and Hunter, the lactoperoxidase method, the gaseous diffusion method and the chloramine-T method. The chloramine-T technique was adapted as the routine method. Ten .mu.g (3.27 nmol) peptide was reacted with 5 mCi of Na125I at a concentration of chloramine-T of 1.3 mmol/l. Synthetic secretin was suitable for labeling for at least 8 mo. when stored as dry matter in N-filled glass ampoules. Purification and separation of labeled from unlabeled hormone was carried out by gel-permeation chromatography on Sephadex G-50 superfine. The labeled preparation had a specific radioactivity of 405 .+-. 33 .mu.Ci/nmol (mean .+-. SEM [standard error of the mean], n = 9) and was usable for 6 wk. 6-Tyrosyl-secretin took up more I compared to porcine synthetic secretin but had lower immunoreactivity with all antisera tested.

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