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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Apr;73(4):1314–1318. doi: 10.1073/pnas.73.4.1314

Fluorometric assay of vasopressin and oxytocin: a general approach to the assay of peptides in tissues.

K A Gruber, S Stein, L Brink, A Radhakrishnan, S Udenfriend
PMCID: PMC430258  PMID: 1063412

Abstract

A fluorometric method for the quantitative assay of vasopressin and oxytocin in individual rat pituitaries has been developed. Acid extracts of pituitaries are freed of amino acids and polyamines by passage over a copper-Sephadex column, and the peptides fraction is then labeled by reaction with fluorescamine. The resulting peptide fluorophors are separated by chromatography on a reverse-phase bonded column. Specificity of the procedure was ascertained by several criteria, including bioassay and amino-acid analysis of the eluted peptide fluorophors. The procedure serves as a model system for the assay of tissue peptides in the picomole range.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Böhlen P., Stein S., Imai K., Udenfriend S. A simplified protein assay with fluorescamine in samples containing interfering material. Anal Biochem. 1974 Apr;58(2):559–562. doi: 10.1016/0003-2697(74)90224-3. [DOI] [PubMed] [Google Scholar]
  2. Böhlen P., Stein S., Stone J., Udenfriend S. Automatic Monitoring of primary amines in preparative column effluents with fluorescamine. Anal Biochem. 1975 Aug;67(2):438–445. doi: 10.1016/0003-2697(75)90316-4. [DOI] [PubMed] [Google Scholar]
  3. FAZAKERLEY S., BEST D. R. SEPARATION OF AMINO ACIDS, AS COPPER CHELATES, FROM AMINO ACID, PROTEIN AND PEPTIDE MIXTURES. Anal Biochem. 1965 Aug;12:290–295. doi: 10.1016/0003-2697(65)90093-x. [DOI] [PubMed] [Google Scholar]
  4. Sachs H., Goodman R., Osinchak J., McKelvy J. Supraoptic neurosecretory neurons of the guinea pig in organ culture. Biosynthesis of vasopressin and neurophysin. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2782–2786. doi: 10.1073/pnas.68.11.2782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Schwabe C., Catlin J. C. Removal of a Fluoram 1-positive impurity from hydrochloric acid. Anal Biochem. 1974 Sep;61(1):302–304. doi: 10.1016/0003-2697(74)90361-3. [DOI] [PubMed] [Google Scholar]
  6. Stein S., Böhlen P., Stone J., Dairman W., Udenfriend S. Amino acid analysis with fluorescamine at the picomole level. Arch Biochem Biophys. 1973 Mar;155(1):202–212. doi: 10.1016/s0003-9861(73)80022-0. [DOI] [PubMed] [Google Scholar]
  7. Stein S., Chang C. H., Böhlen P., Imai K., Udenfriend S. Amino acid analysis with fluorescamine of stained protein bands from polyacrylamide gels. Anal Biochem. 1974 Jul;60(1):272–277. doi: 10.1016/0003-2697(74)90154-7. [DOI] [PubMed] [Google Scholar]
  8. Udenfriend S., Stein S., Böhlen P., Dairman W., Leimgruber W., Weigele M. Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range. Science. 1972 Nov 24;178(4063):871–872. doi: 10.1126/science.178.4063.871. [DOI] [PubMed] [Google Scholar]
  9. Weigele M., De Bernardo S., Leimgruber W. Fluorescent labeling of proteins. A new methodology. Biochem Biophys Res Commun. 1973 Oct 1;54(3):899–906. doi: 10.1016/0006-291x(73)90779-1. [DOI] [PubMed] [Google Scholar]

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