ON-TARGETplus Human GAPDH (2597) siRNA - Individual

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 10^5 cells/well	siRNA concentration-100 nM Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Replace medium after 4h and reincubate cells for 44h at 37°C.

Upstream tips
Seed 10^5 cells/well
Protocol tips
siRNA concentration-100 nM Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Replace medium after 4h and reincubate cells for 44h at 37°C.

Publication protocol

Cells were seeded on 24-well plates (105 cells/well) and cultured overnight to ∼70% confluency. siRNA-polyplexes (corresponding to 100 nM siRNA) were added to the cells in serum-free medium (HBSS:HEPES pH 7.4) and incubated for up to 4 h. Samples were then removed and replaced with fresh culture medium and cells cultured for 44 h. GAPDH activity was analysed using the KDalert GAPDH kit (Ambion, USA) following the supplier’s protocol, as a commonly used knock-down assay.

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Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Human GAPDH (2597) siRNA - Individual below.

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