Development of lanthanide-labeled human INSL3 as an alternative probe to radioactively labeled INSL3 for use in bioassays.
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.
Howard Florey Institute, School of Chemistry, University of Melbourne, Victoria 3010, Australia.
Fazel
F
Shabanpoor
Richard A
RA
Hughes
Ross A D
RA
Bathgate
Frances
F
Separovic
John D
JD
Wade
D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007328:Insulin / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D028581:Lanthanoid Series Elements / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism
D007328:Insulin; D028581:Lanthanoid Series Elements; C084797:Leydig insulin-like protein; D011506:Proteins; C472867:RXFP2 protein, human; D043562:Receptors, G-Protein-Coupled