Silencer® Select GLO-1 siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 2 × 10^5 cells	siRNA concentration-5 nM Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Incubate cells for 1 day at 37°C and change medium. Re-incbaute for another 48h

Upstream tips
Seed 2 × 10^5 cells
Protocol tips
siRNA concentration-5 nM Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Incubate cells for 1 day at 37°C and change medium. Re-incbaute for another 48h

Publication protocol

2 × 105 cells were transfected with 5 nM siRNA GLO1-siRNA (Silencer Select Validated siRNA ID s5824, Ambion Applied Biosystems, USA: sense: 5′-GGCUUAUGAGGAUAAAAAUtt-3′, antisense 5′-AUUUUUAUCCUCAUAAGCCaa-3′) or with a non-target siRNA (NT-siRNA, Silencer Select Negative Control No. 1, Cat. No. 4390849, Ambion Applied Biosystems, USA) using Lipofectamine2000 transfection reagent (Invitrogen, Carlsbad, USA). 50 μM of siRNA and 3 μl of transfection reagent were diluted with Opti-MEM (Invitrogen), and incubated for 20 min at room temperature, and then added to the cells. 1.5 ml transfection medium (EGM-2 with 10% FBS, but without antibiotics) were added. 24 h after transfection, medium was replaced with fresh complete medium containing 25 mM glucose if indicated. Gene expression assays were performed 48 h after transfection and protein expression analysis was done 72 h after transfection, respectively.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Silencer® Select GLO-1 siRNA below.

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