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Development of Lanthanide‐Labeled Human INSL3 as an Alternative Probe to Radioactively Labeled INSL3 for Use in Bioassays
Author(s) -
Shabanpoor Fazel,
Hughes Richard A.,
Bathgate Ross A. D.,
Separovic Frances,
Wade John D.
Publication year - 2009
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2009.03839.x
Subject(s) - europium , chemistry , bioassay , peptide , chelation , lanthanide , chromatography , combinatorial chemistry , fluorescence , biochemistry , biology , organic chemistry , ion , genetics , physics , quantum mechanics
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.