Protocol for measuring dioxin-like activity in environmental sample using in vitro reporter gene dr-luc assays.
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Schipper, Cor A.
Leonards, Pim E. G.
Klamer, Hans J. C.
Thomas, Kevin V.
Vethaak, Andre D.
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Guidelines are given for the quantification of the dioxin-like activities of contaminants in sediment, biota and water samples using the DR-Luc reporter gene bioassay. Dioxins and dioxin-like compounds demonstrate high affinity binding to the Aryl hydrocarbon Receptor (AhR). Ah-R is a ligand-activated transcription factor and mediates most, if not all, of the toxic responses of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (PCBs), and polybrominated biphenyls (PBBs). The DR-Luc bioassay, or DR-CALUX® (Dioxin Response Chemical Activated LUciferase gene eXpression), utilizes a recombinant rat hepatoma H4IIE cell line with a stably integrated AhR- responsive luciferase reporter gene. Exposure of this bioassay to extracts containing dioxin-like compounds induces the enzyme luciferase in a time, dose, and chemical specific manner. Cells are cultured in the laboratory and transferred to 96-well plates. Luciferase activity.....
PublisherInternational Council for the Exploration of the Sea (ICES)
Series;NrICES Techniques in Marine Environmental Sciences; 55
Sustainable Development Goals (SDG)14.1
Best Practice TypeStandard Operating Procedure
CitationSchipper, C.A.; Leonards, P.E.G.; Klamer, H.J.C., Thomas, K.V. and Vethaak, A.D. (2013) Protocol for measuring dioxin-like activity in environmental samples using in vitro reporter gene DR-Luc assays. ICES Techniques in Marine Environmental Sciences, No. 55, 21pp. DOI: http://dx.doi.org/10.25607/OBP-264