FAK siRNA (h)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 2 × 10^5	Add the siRNA duplex solution (Solution A) directly to the dilute Transfection Reagent (Solution B) using a pipette. Mix gently by pipetting the solution up and down and incubate the mixture 15-45 minutes at room temperature. Wash the cells once with 2 ml of siRNA Transfection Medium. Add siRNA Transfection Reagent mixture (Solution A + Solution B) to cells and incubate the cells 5-7 hours at 37° C in a CO2 incubator and replace medium. Incubate the cells for an additional 18-24 hours.

Upstream tips
Seed 2 × 10^5
Protocol tips
Add the siRNA duplex solution (Solution A) directly to the dilute Transfection Reagent (Solution B) using a pipette. Mix gently by pipetting the solution up and down and incubate the mixture 15-45 minutes at room temperature. Wash the cells once with 2 ml of siRNA Transfection Medium. Add siRNA Transfection Reagent mixture (Solution A + Solution B) to cells and incubate the cells 5-7 hours at 37° C in a CO2 incubator and replace medium. Incubate the cells for an additional 18-24 hours.

Publication protocol

For silencing specific gene expression, U251 glioma cells were treated with FAK siRNA, Pyk2 siRNA and integrin β3 siRNA (Santa Cruz Biotechnology, USA). Briefly, 2 × 105 U251 glioma cells were seeded into six well plate with 2 ml antibiotic-free normal growth medium containing FBS. Transfection of FAK siRNA, Pyk2 siRNA, integrin β3 siRNA or control siRNA was performed according to the manufacture’s protocol. U251 glioma cells transfected with siRNA for 24 h were then exposed to hypoxia.

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Manufacturer protocol

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

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