Fluorescence-based enzymatic assay by capillary electrophoresis laser-induced fluorescence detection for the determination of a few β-galactosidase molecules

Doug Craig, Edgar A. Arriaga, Peter Banks, Yanni Zhang, Annika Renborg, Monica M. Palcic, Normal J. Dovichi

Research output: Contribution to journalArticlepeer-review

48 Scopus citations

Abstract

β-Galactosidase can be assayed by monitoring the generation of the fluorescent products, fluorescein-mono-β-D-galactopyranoside and fluorescein, when the fluorogenic substrate fluorescein-di-β-D-galactopyranoside is used. We have used capillary electrophoresis with ultrasensitive laser-induced fluorescence detection to monitor the formation of the fluorescent products of off-column enzymatic reactions. By analyzing as little as 40 pl of the enzymatic mixture we obtain limits of detection of 6.5 × 10-14 M β-galactosidase or 1.6 molecules based on the detection of fluorescein-mono-β-D-galactopyranoside.

Original languageEnglish (US)
Pages (from-to)147-153
Number of pages7
JournalAnalytical Biochemistry
Volume226
Issue number1
DOIs
StatePublished - Mar 1995

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