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Title of Journal: Sci China Chem

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Abbravation: Science China Chemistry

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SP Science China Press

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DOI

10.1002/ardp.18450920116

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1862-2771

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Enhancing the response of CALUX and CAFLUX cell bi

Authors: Bin Zhao David S Baston Elaine Khan Claudio Sorrentino Michael S Denison
Publish Date: 2010/06/02
Volume: 53, Issue: 5, Pages: 1010-1016
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Abstract

Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications Over the past 10 years reporter gene assays have been widely accepted and used for analysis of 2378tetrachlorodibenzopdioxin and related dioxinlike compounds in various types of matrices such as biological environmental food and feed samples given that highresolution instrumental analysis techniques are impractical for largescale screening analysis The most sensitive cellbased reporter gene bioassay systems developed are the mechanismbased CALUX Chemically Activated Luciferase Expression and CAFLUX Chemically Activated Fluorescent Expression bioassays which utilize recombinant cell lines containing stably transfected dioxin AhRresponsive firefly luciferase or enhanced green fluorescent protein EGFP reporter genes respectively While the current CALUX and CAFLUX bioassays are very sensitive increasing their lower limit of sensitivity magnitude of response and dynamic range for chemical detection would significantly increase their utility particularly for those samples that contain low levels of dioxinlike HAHs ie serum In this study we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays

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