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

2.1. Cell culture and transfection
Human dopaminergic neuroblastoma cells (SH-SY5Y, ATCC, Manassas, VA, USA) were grown in Dulbecco's modified Eagle medium
(DMEM)/F-12 medium (Nacalai Tesque, Kyoto, Japan) supplemented
with 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA),
50 U/ml penicillin (Nacalai Tesque, Kyoto, Japan), and 50 μg/mL streptomycin (Nacalai Tesque, Kyoto, Japan). The growth conditions were
maintained at 37 °C in a humidified environment containing 95% air
and 5% CO2. The pEGFP-N1/FtMt plasmid was established in a previous
study (Wang et al., 2011) and this plasmid does not express green
fluorescent protein (GFP) because EGFP is located downstream of the
multiple cloning site. SH-SY5Y cells were transiently transfected with
pEGFP-N1 (empty vector; Clontech Laboratories, Inc., Palo Alto, CA,
USA) as a control and pEGFP-N1/FtMt using FuGENE HD transfection
reagent (Promega, Madison, WI, USA) according to the manufacturer's
instructions. The experiment was continued for 24 or 48 h after transfection. All of the experimental procedures employed in this study
were approved by the Gene Recombination Experiment Safety Committee of Shiga University of Medical Science.

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Manufacturer protocol

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

Download manufacturer protocol

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