siRNA / miRNA gene silencing Human - PC3 (human prostate cancer cell line) STEAP2

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

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STEAP2 metalloreductase

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	PC3 cells purchased from American Type Culture Collection (ATCC, LGC Standard) were maintained in RPMI 1640 media (Gibco, Life Technologies, UK) supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, and 100 U/ml penicillin/streptomycin (Gibco, Life Technologies, UK) at 37 °C and 5% CO2. STEAP2 expression was reduced using siRNA S48991, a non-specific siRNA (4390843) was used as a control (Life Technologies, UK) and Lipofectamine RNAiMAX (Life Technologies, UK) was used according to manufacturer’s instructions. PC3 cells were incubated with the siRNA complex for 72 h.

Protocol tips
PC3 cells purchased from American Type Culture Collection (ATCC, LGC Standard) were maintained in RPMI 1640 media (Gibco, Life Technologies, UK) supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, and 100 U/ml penicillin/streptomycin (Gibco, Life Technologies, UK) at 37 °C and 5% CO2. STEAP2 expression was reduced using siRNA S48991, a non-specific siRNA (4390843) was used as a control (Life Technologies, UK) and Lipofectamine RNAiMAX (Life Technologies, UK) was used according to manufacturer’s instructions. PC3 cells were incubated with the siRNA complex for 72 h.

Protocol tips

PC3 cells purchased from American Type Culture Collection (ATCC, LGC Standard) were maintained in RPMI 1640 media (Gibco, Life Technologies, UK) supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, and 100 U/ml penicillin/streptomycin (Gibco, Life Technologies, UK) at 37 °C and 5% CO2. STEAP2 expression was reduced using siRNA S48991, a non-specific siRNA (4390843) was used as a control (Life Technologies, UK) and Lipofectamine RNAiMAX (Life Technologies, UK) was used according to manufacturer’s instructions. PC3 cells were incubated with the siRNA complex for 72 h.

Manufacturer protocol Publication protocol 1 Relevant paper

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