A new antiserum with conformational specificity for LHRH: Usefulness for radioimmunoassay and immunocytochemistry

William E. Ellinwood, Oline K. Ronnekleiv, Martin J. Kelly, John A. Resko

Research output: Contribution to journalArticlepeer-review

98 Scopus citations

Abstract

We have produced and characterized a new high titer, highly specific antiserum for luteinizing hormone-releasing hormone (LHRH), and demonstrated its usefulness for radioimmunoassay (RIA) and immunocytochemistry. The antiserum can be used at a final dilution of 1:500,000 to 1:600,000 for RIAs with a sensitivity of 0.2 pg/tube. Both the amino and carboxy terminal ends of the LHRH molecule are required for antibody recognition, and the antigenic determinant appears to be part of a three-dimensional structure of LHRH. Fragments of LHRH and other brain peptides are not recognized by the antiserum. Using immunocytochemical techniques, we have localized LHRH-containing neurons in the medial basal hypothalamus of the rhesus monkey, guinea pig, and rat. Staining of LHRH fibers and cell bodies was eliminated by preabsorbtion of the immune serum with synthetic LHRH. This antiserum should be useful in studies that require quantification of very low amounts of LHRH and in studies that require correlation between immunocytochemical localization and tissue content or secretion of LHRH.

Original languageEnglish (US)
Pages (from-to)45-52
Number of pages8
JournalPeptides
Volume6
Issue number1
DOIs
StatePublished - Jan 2 1985

Keywords

  • Immunoctyochemistry
  • LHRH
  • Radioimmunoassay

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Endocrinology
  • Cellular and Molecular Neuroscience

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