TRAF6 siRNA (h)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1×10^6 cells/well	Add diluted siRNA to diluted Lipofectamine Reagent   Incubate for 20 min at RT and add to cells.  Replace medium after 6h and re-incubate cells for 2 days at 37°C.

Upstream tips
Seed 1×10^6 cells/well
Protocol tips
Add diluted siRNA to diluted Lipofectamine Reagent   Incubate for 20 min at RT and add to cells.  Replace medium after 6h and re-incubate cells for 2 days at 37°C.

Publication protocol

Log-phase SPC-A1 cells were seeded onto 6-well plates (Corning, Inc., Corning, NY, USA) at a density of 1×106 cells/well, and incubated at 37°C for 24 h. Four micrograms of the specific siRNA targeting TRAF6 or control siRNA (both from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) was added to 250 µl of serum-free DMEM and mixed evenly, while 10 µl of Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) was added to another 250 µl of serum-free DMEM, gently mixed evenly and placed at room temperature for 5 min. The siRNA and Lipofectamine 2000 solutions were mixed evenly, and placed at room temperature for 20 min. Then, the mixture was transferred to 6-well plates, mixed evenly and gently and incubated at 37°C. Following a 6-h transfection, the cell culture solution was changed, and to each well h 2 ml medium was added. After 48 h of transfection, the cells were harvested for the subsequent experiments, and non-transfected cells served as blank controls. Transfection efficiency was verified using western blot assay.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for TRAF6 siRNA (h) below.

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