Lipofectamine® 2000 Transfection Reagent

Protocol tips

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	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Transfection and silencing
For transient transfections of Tet-21/N and SH-SY5Y, cells cultured on 100-mm dishes were transfected with 12 μg of SESN2 plasmid, GFP-mRFP-LC3 (kind gift from Dr A Fraldi, Tigem Institute) construct or empty vector using Lipofectamine 2000. For LSD1 knockdown, 50 or 100 nm siRNA targeting LSD1 or LSD2 (GE Dharmacon, Lafayette, CO, USA) or scramble were transfected in Tet-21/N cells using a MicroPorator Digital Bio Technology(Waltham, MA, USA), according to the recently described protocol.41 Briefly, 2x106 cells were collected by trypsin/EDTA digestion, washed once with calcium and magnesium-free phosphate-buffered saline (PBS) and resuspended in 100 μl of resuspension buffer, mixed with siRNA or scramble and electroporated as described.25 The efficiency of LSD1 siRNA knockdown was assayed 48 h after transfection by western blotting. For silencing assays, 45 nm siRNA targeting SESN2 (GE Dharmacon) or scramble were transfected in SH-SY5Y cells using Viromer Green; Cells were collected for analysis 48 h after transfection.

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Manufacturer protocol

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

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