Fluoroprofile, a fluorescence-based assay for rapid and sensitive quantitation of proteins in solution

James A. Mackintosh, Duncan A. Veal, Peter Karuso*

*Corresponding author for this work

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The development of a sensitive fluorescence-based assay for the quantitative determination of protein concentration is described. The assay is based on the natural product epicocconone, which produces a large increase in fluorescence quantum yield upon binding to detergent-coated proteins in solution. There is a concomitant shift in the emission maximum from 520 to 605 nm after binding, which results in low background signal allowing a linear dynamic range of 40 ng/mL to 200 μg/mL for most proteins. There is little protein-to-protein variation except for iron-containing proteins and the assay can be used so that it is tolerant of chemicals commonly used in 2-D sample buffers. The assay is more sensitive than standard absorption assays such as the Bradford and Lowry assays, and has a greater dynamic range and sensitivity than other fluorescent assays.

Original languageEnglish
Pages (from-to)4673-4677
Number of pages5
JournalProteomics
Volume5
Issue number18
DOIs
Publication statusPublished - Dec 2005

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