HIF-1α siRNA (r)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 2×10^5 cells per well in a six-well plate	Add solution -A and solution-B and incubate the mixture 15-45 minutes at room temperature. Add the mixture to cells and incubate the cells 5-7 hours at 37° C in a CO2 incubator and later add normal growth medium and incubate for an additional 18-24 hours.

Upstream tips
Seed 2×10^5 cells per well in a six-well plate
Protocol tips
Add solution -A and solution-B and incubate the mixture 15-45 minutes at room temperature. Add the mixture to cells and incubate the cells 5-7 hours at 37° C in a CO2 incubator and later add normal growth medium and incubate for an additional 18-24 hours.

Publication protocol

rBMECs were seeded at a density of 2×105 cells per well in a six-well plate and were grown in antibiotic-free medium until they reached 60%–80% confluence. They were then subjected to gene silencing using small-interfering-RNA (siRNA) targeting HIF-1α or an HIF-1α scrambled control and a transfection reagent (sc-45919, sc-37007 and sc-29528, Santa Cruz) according to the manufacturer's instructions. The transfection efficiency was evaluated using Western blot analysis of HIF-1α.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for HIF-1α siRNA (r) below.

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