期刊
JOURNAL OF BIOMOLECULAR SCREENING
卷 8, 期 4, 页码 410-420出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057103256319
关键词
fluorescence polarization; high-throughput screening; CyDye-labeled nonpeptide and peptide ligands; miniaturization of FP receptor assays
资金
- NIMH NIH HHS [N01MH30003] Funding Source: Medline
Fluorescence polarization (FP) is an established technique for the study of biological interactions and is frequently used in the high-throughput screening (HTS) of potential new drug targets. This work describes the miniaturization of FP receptor assays to 1536-well formats for use in HTS. The FP assays were initially developed in 384-well microplates using CyDye-labeled nonpeptide and peptide ligands. Receptor expression levels varied from similar to1 to 10 pmols receptor per mg protein, and ligand concentrations were in the 0.5- to 1.0-nM range. The FP assays were successfully miniaturized to 1536-well formats using Cy3B-labeled ligands, significantly reducing reagent consumption, particularly the receptor source, without compromising assay reliability. Z' factor values determined for the FP receptor assays in both 384- and 1536-well formats were found to be >0.5, indicating the assays to be robust, reliable, and suitable for HTS purposes.
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