Nrf2 siRNA (r)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add Solution A to Solution B and incubate for 15-45 minutes at room temperature. Add this mixture to cells and incubate for 5-7 hours at 37° C in a CO2 incubator. Add 1 ml of normal growth medium containing 2 times the normal serum and antibiotics concentration (2x normal growth medium) without removing the transfection mixture. Incubate the cells for an additional 18-24 hours.

Protocol tips
Add Solution A to Solution B and incubate for 15-45 minutes at room temperature. Add this mixture to cells and incubate for 5-7 hours at 37° C in a CO2 incubator. Add 1 ml of normal growth medium containing 2 times the normal serum and antibiotics concentration (2x normal growth medium) without removing the transfection mixture. Incubate the cells for an additional 18-24 hours.

Publication protocol

Astrocytes (DIV10–14) were transfected with 20–30 pmol of scrambled or nuclear factor erythroid 2-related factor 2 (Nrf2)-targeting siRNA (Santa Cruz, catalog numbers SC-37007 and SC-156128, respectively) using Nucleofector II (Amaxa) according to the manufacturer’s protocol. Based on separate experiments with GFP expression plasmid pmaxGFP (Lonza), astrocyte transfection efficiency using this approach is estimated at 70–80%

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Manufacturer protocol

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