FuGENE® HD Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Always bring FuGENE HD to room temperature and mix well prior to use.

Upstream tips
Always bring FuGENE HD to room temperature and mix well prior to use.

Publication protocol

Transient Transfection and Luciferase Activity Assay
MCF-7 cells and A2780 cells were seeded into 100-mm cell culture dishes and reached 70–80% confluence 24 h after plating. The cells were then transfected with the pGL3-HRE-luciferase plasmid using FuGENE HD transfection reagent (Roche Applied Science) as described previously (16). The next day, cells were split into 24-well plates at a density of 2 × 105 cells/well. 48 h after transfection, cells were treated for 4 h with clofibrate, fenofibrate, troglitazone, and other reagents at the indicated concentrations and durations and then placed into a hypoxia chamber (1% O2 and 5% CO2; ProOx model C21, Biospherix, Ltd., Lacona, NY) for 16 h. Cell lysate was prepared, and luciferase activity was assayed using the Dual-Luciferase reporter kit as described (16). The firefly luciferase activity was normalized for the amount of protein used for luciferase activity assay for each sample. The data are expressed as percentages of luciferase activity detected in untreated cells.

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Manufacturer protocol

Download the product protocol from Promega for FuGENE® HD Transfection Reagent below.

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Videos

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