Book Chapter
Development of lanthanide-labeled human INSL3 as an alternative probe to radioactively labeled INSL3 for use in bioassays
F Shabanpoor, RA Hughes, RAD Bathgate, F Separovic, JD Wade
Annals of the New York Academy of Sciences | WILEY-BLACKWELL | Published : 2009
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. © 2009 New York Academy of Sciences.
Grants
Awarded by NHMRC
Funding Acknowledgements
We thank Sharon Layfield for carrying Out the Cell Culture and Tania Ferrero for help with binding assays. This work was funded by NHMRC project grant 350245 to JDW, RADB, and RAH.