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Mutat Res. 2002 Jan 15;513(1-2):169-82.

Direct influence of S9 liver homogenate on fluorescence signals: impact on practical applications in a bacterial genotoxicity assay.

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1
Department of Biological Technologies, F. Hoffmann-La Roche Ltd., Pharma Division, CH-4002 Basel, Switzerland. juerg.dreier@basilapharma.com

Abstract

Assays based on the bacterial SOS-response offer the possibility of automatization of genotoxicity testing for screening of large compound libraries. While existing assays use colorimetric detection or luminescence read-out, we describe here the use of a fluorescence-based system to achieve high sensitivity of detection required for assay miniaturization. Three commonly used fluorophores--fluorescein, DDAO and resorufin--are evaluated. Experimental evidence is given that S9 liver homogenate contains a heat-labile, reversible fluorophore-binding activity and therefore, significantly reduces fluorescence intensities. We have worked out simple solutions to overcome the S9 related interference in order to be able to establish a robust bacterial genotoxicity assay.

PMID:
11719102
[Indexed for MEDLINE]
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