Development of lanthanide-labeled human INSL3 as an alternative probe to radioactively labeled INSL3 for use in bioassays

Ann N Y Acad Sci. 2009 Apr;1160:87-90. doi: 10.1111/j.1749-6632.2009.03839.x.

Abstract

The use of radioisotope-labeled peptides and proteins in bioassays is diminishing as a consequence of the emergence of fluorescent lanthanide-labeled probes. We describe a simple, single-step labeling of synthetic human insulin-like peptide 3 (INSL3) with a commercially available activated europium chelator. The labeled peptide was then used for the development of a robust and highly reproducible receptor-binding assay which is amenable to automated high-throughput screening of ligands. Europium-labeled INSL3 showed a similar binding profile to radioactively labeled INSL3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay / methods*
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Humans
  • Insulin / chemical synthesis
  • Insulin / chemistry*
  • Insulin / metabolism
  • Lanthanoid Series Elements / chemical synthesis
  • Lanthanoid Series Elements / chemistry*
  • Lanthanoid Series Elements / metabolism
  • Proteins / chemical synthesis
  • Proteins / chemistry*
  • Proteins / metabolism
  • Receptors, G-Protein-Coupled / metabolism

Substances

  • Insulin
  • Lanthanoid Series Elements
  • Leydig insulin-like protein
  • Proteins
  • RXFP2 protein, human
  • Receptors, G-Protein-Coupled