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

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

Research output: Contribution to journalArticleResearchpeer-review

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.

LanguageEnglish
Pages4673-4677
Number of pages5
JournalProteomics
Volume5
Issue number18
DOIs
Publication statusPublished - Dec 2005

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Assays
Fluorescence
Proteins
Biological Products
Detergents
Quantum yield
Buffers
Iron

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Mackintosh, James A. ; Veal, Duncan A. ; Karuso, Peter. / Fluoroprofile, a fluorescence-based assay for rapid and sensitive quantitation of proteins in solution. In: Proteomics. 2005 ; Vol. 5, No. 18. pp. 4673-4677.
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Fluoroprofile, a fluorescence-based assay for rapid and sensitive quantitation of proteins in solution. / Mackintosh, James A.; Veal, Duncan A.; Karuso, Peter.

In: Proteomics, Vol. 5, No. 18, 12.2005, p. 4673-4677.

Research output: Contribution to journalArticleResearchpeer-review

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