Cxcr4 siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Final concentartion of siRNA- 100 pmol per plate. Incubate Lipofectamine reagent:siRNA mixture for 5 min and add to cells. Treat cells with Sdf-1 (10 ng/μl) after 24 h. Incubate cells for another 48h at 37°C and quantify.

Protocol tips
Final concentartion of siRNA- 100 pmol per plate. Incubate Lipofectamine reagent:siRNA mixture for 5 min and add to cells. Treat cells with Sdf-1 (10 ng/μl) after 24 h. Incubate cells for another 48h at 37°C and quantify.

Publication protocol

C2C12 or ESCs were plated on plates covered with Matrigel Matrix Growth Factor Reduced (BD Biosciences). After reaching 30 to 40% confluence the cells were transfected with Silencer Select Pre-designed siRNA (Life Technologies) complementary to mRNAs encoding either CXCR4 9 (ID:s64091) or CXCR7 (ID:s64124). Appropriate, recommended negative control siRNA was used. siRNA duplexes were diluted in DMEM to reach the concentration of 100 pmol per plate and incubated with Lipofectamine RNAiMAX (Life Technologies), according to the manufacturer’s instructions. After 24 hours the cells were treated with Sdf-1 (10 ng/μl). Next, cells were collected 48 hours post Sdf-1 treatment and processed either for mRNA isolation followed by quantitative RT-PCR, immunolocalisation, or western blotting. The efficiency of CXCR4 or CXCR7 downregulation was tested by quantitative RT-PCR and immunocytochemistry.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Cxcr4 siRNA below.

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