Development of a ubiquitin transfer assay for high throughput screening by fluorescence resonance energy transfer.
Authors:Boisclair MD, McClure C, Josiah S, Glass S, Bottomley S, Kamerkar S, Hemmilä I
Journal:J Biomol Screen
PubMed ID:11080690
'An assay based on fluorescence resonance energy transfer (FRET) has been developed to screen for ubiquitination inhibitors. The assay measures the transfer of ubiquitin from Ubc4 to HECT protein Rsc 1083. Secondary reagents (streptavidin and antibody to glutathione-S-transferase [GST]), pre-labeled with fluorophores (europium chelate, Eu(3+), and allophycocyanin [APC]), are noncovalently ... More