FuGENE® 6 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 transfections and luciferase assays
Transient transfection analysis was performed as previously described []. Constructs containing 216 bp of the rat elastin promoter driving luciferase, referred to as 216-luc [] and a nuclear factor κ B (NFκB) reporter construct driving luciferase expression, referred to as NFκB-luc [] were used. Cultures were co-transfected with the experimental construct and a pRL-CMV- construct (Promega, Madison, WI) (the latter for normalization of transfection efficiency). To achieve optimal efficiency, transfections were performed using FuGene 6 (Roche, Indianapolis; following the manufacturer’s instructions) when SMCs were approximately 70% confluent. Twenty-four hours after transfection, media were removed and the cells were cultured under experimental conditions as indicated. At the time of harvest, media were removed, cells were washed twice with cold HBSS and placed at -80°C for at least 24 hours. To determine luciferase and activities, the Dual-luciferase Reporter Assay (Promega) kit was used as per manufacturer’s instructions. Data are expressed as luciferase/ ± SD.

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Papers

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Manufacturer protocol

Download the product protocol from Promega for FuGENE® 6 Transfection Reagent below.

Download manufacturer protocol

Videos

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