Wu, Meng and Lin, Zhihong and Wolfbeis, Otto S. (2003) Determination of the activity of catalase using a europium(III)-tetracycline-derived fluorescent substrate. ANALYTICAL BIOCHEMISTRY, 320 (1). pp. 129-135. ISSN 0003-2697
Full text not available from this repository.Abstract
A one-step method is described for the fluorometric determination of the activity of the enzyme catalase (EC 1.11.1.6.), based on the finding that H2O2 in the europium (III)-tetracycline-hydrogen peroxide system is consumed by catalase. This is accompanied by a large decrease in both fluorescence intensity and decay time. The limit of detection (LOD; at S/N=3) for catalase at 30degreesC for a 10-min kinetic assay is 1.0 unit/mL, with a linear range from 1.0 to 10 unit/mL. At an incubation time of 30 min at 37degreesC for a one-point assay, the LOD is 0.046 unit/mL, with a linear range from 46 to 400 munit/mL. The assay was performed on microtiterplates and is fully compatible with existing plate readers. It is a one-step, simple, and sensitive method suitable for both continuous kinetic and one-point detections, does not require the addition of other substrates, and works best at neutral pH (with an optimum at pH 6.9). The reagent has the typical spectral features of a europium-ligand complex including a large Stokes shift (210 nm), a red line-like emission (centered at 616 nm), and a decay time in the microsecond domain. It is also the first europium-based probe that is compatible with the 405-nm diode laser. In summary, the new assay provides distinct advantages over direct ultraviolet detection and over the two-reagent (peroxidase) method. (C) 2003 Elsevier Science (USA). All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HYDROGEN-PEROXIDE; catalase; europium; tetracycline; hydrogen peroxide |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, formerly Prof. Wolfbeis) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 29 Jul 2021 10:37 |
| Last Modified: | 29 Jul 2021 10:37 |
| URI: | https://pred.uni-regensburg.de/id/eprint/38616 |
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