Ccr2 siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Dilute Lipofectamine 2000 in DMEM/F12. Combine the above mixture with 0.01 to 0.20 µg siRNA Incubate for 15min at RT. Incubate cells for 1 day at 37°C	Analyze transfected cells with western blot analysis.

Protocol tips
Dilute Lipofectamine 2000 in DMEM/F12. Combine the above mixture with 0.01 to 0.20 µg siRNA Incubate for 15min at RT. Incubate cells for 1 day at 37°C
Downstream tips
Analyze transfected cells with western blot analysis.

Publication protocol

The method for preparing and transfecting siRNA has been described in detail previously [26]. Briefly, CCR2 siRNAs were purchased from Ambion (Austin, TX) and complexed with Lipofectamine 2000 (Invitrogen, Karlsruhe, Germany) in six-well plates according to the manufacturer’s instructions. Two microliters Lipofectamine 2000 was diluted in 50 µl Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F-12 (DMEM/F12; Sigma, St Louis, MO) and combined with 0.01 to 0.20 µg siRNA after a 15-min incubation at room temperature. The transfection was continued for 24 h at room temperature. The knockdown of CCR2 in microglia was determined with western blot analysis.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Ccr2 siRNA below.

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