Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Overexpression of αB-crystallin
Plasmid pcDNA3 rat-αBC was provided by Dr. J-K. Lee (Inha University, Incheon, South Korea). Chondrocyte were seeded in six-well plates (10 cells/well) and were transiently transfected with each construct using Lipofectamine 2000, according to the manufacturer's instructions. Briefly, 2μg of plasmid DNA was mixed with 6μl of Lipofectamine 2000 and then incubated with Opti-MEM. The plasmid DNA–Lipofectamine 2000 complex was added to the cells, and the mixture was further incubated for 4 h at 37°C. After incubation, the medium was replaced with 2 ml of growth medium, and the cells were maintained for an additional 20 h. Twenty-four hours after transfection, the plasmid DNA transfection medium was removed, and each well was washed with PBS solution. Cells were further exposed new fresh medium for 24 h. V, cells tranfected with pcDNA3 vector.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine® 2000 Transfection Reagent below.

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