X-tremeGENE™ siRNA Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add diluted siRNA to transfection reagent within 5 min. Incubate for 15-20 minutes at room temperature and add to cells.

Protocol tips
Add diluted siRNA to transfection reagent within 5 min. Incubate for 15-20 minutes at room temperature and add to cells.

Publication protocol

Transfections were performed according to the manufacturer's instructions. To knock down Hsc70, synthesized duplex siRNAs were produced by GenePharma Co. Ltd (Shanghai, China). Double‐stranded siRNA was designed (sense, 5′‐GGCCAGUAUUGAGAUUGAUTT‐3′; antisense, 5′‐AUCAAUCUCAAUACUGGCCTT‐3′) with two thymidine residues introduced at the 3′ end. A non‐specific control siRNA (sense, 5′‐UUCUCCGAACGUGUCACGU‐3′; antisense 5′‐ACGUGACACGUUCGGAGAA‐3′) was also produced. AR42J cells were grown to 50% confluence in six‐well plates and transfected with the siRNAs in serum‐free medium without antibiotic supplements using the X‐tremeGENE siRNA transfection reagent (Roche Applied Science, Shanghai, China). Silencing of protein expression was confirmed by subsequent western blot analysis. After transfection, the cells were treated with cerulein and H2 under the aforementioned conditions and then harvested for further experiments.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for X-tremeGENE™ siRNA Transfection Reagent below.

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