Enzymic iodination of the histidyl residue of secretin: A radioimmunoassay of the hormone

Holohan, K.Niall; Murphy, Richard F.; Flanagan, R; Buchanan, K. D.; Elmore, D. T.

doi:10.1016/0005-2795(73)90190-6

Cited by 52 publications

(17 citation statements)

References 15 publications

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“…Three further boosters were given at intervals of 4 months using similar techniques but with haemocyanin from the keyhole limpet (Sigma Chemical Co, Poole, Dorset) as the protein carrier instead of BSA. 125I-Labelled GIP was prepared using a modification of an enzymatic iodination method (Holohan, Murphy, Flanagan, Buchanan & Elmore, 1973) carried out at room temperature. Two nmol GIP were dissolved in 50 pi 005 mol sodium acetate/1, pH 50, followed by rapid serial additions of approximately 0-5 nmol Na125I (37 MBq; IMS 30,555 MBq/pg I; The Radiochemical Centre, Amersham), 22 pmol lactoperoxidase derived from milk (Sigma Chemical Co) and I -8 nmol H202/1, the latter two in 10 pi acetate buffer.…”

Section: Methodsmentioning

confidence: 99%

A Radioimmunoassay of Gastric Inhibitory Polypeptide in Human Plasma

Sarson¹,

Bryant²,

Bloom³

1980

Journal of Endocrinology

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A sensitive radioimmunoassay for the measurement of human gastric inhibitory polypeptide (GIP), using pure porcine GIP, has been developed. Cross-reactivity of the antiserum with all available mammalian gut peptide preparations was negligible with the exception of glucagon when it was approximately 1%. Two major molecular forms of GIP were detectable in plasma and tissue extracts, one of large molecular size and the other corresponding to the elution coefficient of pure porcine standard. Concentrations of GIP in plasma from 50 normal subjects after overnight fasting were 9 +/- 1.0 (S.E.M.) pmol/l rising to a peak of 34 +/- 2.8 pmol/l following the ingestion of a small mixed test meal. Ingestion of glucose or fat resulted in a similar rise of plasma GIP, whereas no change was observed after the ingestion of protein.

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Section: Methodsmentioning

confidence: 99%

A Radioimmunoassay of Gastric Inhibitory Polypeptide in Human Plasma

Sarson¹,

Bryant²,

Bloom³

1980

Journal of Endocrinology

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“…Secretin was labelled with 1125 (Radiochemical Centre, Amersham) by the method of Holohan [9] and antibodies were raised to both synthetic secretin and pork natural secretin [10] (kindly donated by Dr. V. Mutt). Two antibodies were used in the studies BB33 raised against synthetic secretin (final titre 1:20,000 to 36,000); and BB101 raised against natural secretin (final titre 1 : 36,000).…”

Section: Methodsmentioning

confidence: 99%

Secretin and insulin: Response to intraduodenal acid

et al. 1976

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Summary.Immunoreactive secretin (IRS) and immunoreactive insulin (IRI) levels were measured in humans and dogs following the intraduodenal instillation of hydrochloric acid. IRS levels rose after acid in both instances, but a concomitant rise in peripheral IRI levels was not noted. Premedication of the humans with Scopolamine prevented a rise of IRS in the human subjects. It is concluded that the endogenous release of IRS alone does not result in increased IRI levels in peripheral blood and that IRS release may be under vagal control.

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“…After 4 months, the animals were each given a booster injection of 40 nmol conjugated neurotensin and the antisera extracted. Radioactively labelled neurotensin was prepared by a modification of the lactoperoxidase iodination method (Holahan, Murphy, Flanagan, Buchanan & Elmore, 1973). This was carried out at room temperature by the serial addition of 100 µ 0-01 M-acetate buffer, pH 5-0, 1 mCi 125I (approx.…”

Section: Introductionmentioning

confidence: 99%