Ccl2 siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA seq:sense 5′-GGCUAAAAGUAGUGAAUGAtt-3′, antisense 5′-UCAUUCACUACUUUUAGCCtt-3′, siRNA concentartion:0.01 to 0.20 μg. Incubate mixture for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C

Protocol tips
siRNA seq:sense 5′-GGCUAAAAGUAGUGAAUGAtt-3′, antisense 5′-UCAUUCACUACUUUUAGCCtt-3′, siRNA concentartion:0.01 to 0.20 μg. Incubate mixture for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C

Publication protocol

MCP-1 siRNA was purchased from Ambion (Austin, TX) and had the following sequences: sense 5′-GGCUAAAAGUAGUGAAUGAtt-3′, antisense 5′-UCAUUCACUACUUUUAGCCtt-3′, and negative control siRNA (sense 5′-CCUACGCCACCAAUUUCGUtt-3′, antisense 5′-ACGAAAUUGGUGGCG UAGGtt-3′). MCP-1 siRNA was complexed with Lipofectamine 2000 (Invitrogen, Karlsruhe, Germany) in 6-well plates in which 7-day-old primary purified retinal neuronal cells were incubated according to the manufacturer's instructions. Then, 2 μL Lipofectamine 2000 was diluted in 50 μL neurobasal medium without supplements and combined with 0.01 to 0.20 μg siRNA after 5-minute incubation at room temperature. The transfection was continued for 24 hours at room temperature. The knockdown of MCP-1 in neuronal cells was determined by Western blot analysis. The process of Western bolt was the same as described above.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Ccl2 siRNA below.

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