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

2.3. Gene transfection
HepG2 cells were seeded at 1.0 × 104 cells in a collagen-coated 96-well microplate and incubated at 37 °C under 5% CO2. The cells were cultured in DMEM containing 10% FBS and 100 U/ml penicillin, and 0.1 mg/ml streptomycin. After 24 h, the cells were transfected with the pDNA at 0.2 μg/ml using the lipoplex or the Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer's instruction. In brief, on the day of transfection, the medium was removed and replaced with fresh medium with or without serum. DA/pDNA and Lipofectamine 2000/pDNA complexes were added into the serum containing and free wells, respectively. After 6 h, the dish transfected with Lipofectamine 2000 was replaced with fresh medium containing serum. After 48 h, the cells were washed with PBS twice adequately and then lysed with a lysis buffer from the luciferase assay kit (Promega). The luciferase activity in a 10 μL aliquot of the cell lysate was measured with a luminescence plate reader (Wallac 1420; Perkin Elmer, Wellesley, MA). The protein concentration of each well lysate was determined by a standard protein assay (Dojindo, Kumamoto, Japan). The luciferase activity in each sample was normalized to the luminescence intensity per microgram of protein.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA transfection Mammalian cells - Immortalized cell lines HepG2 using Lipofectamine® 2000 Transfection Reagent from Thermo Fisher Scientific.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA transfection Mammalian cells - Immortalized cell lines HepG2 using Lipofectamine® 2000 Transfection Reagent from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.