siGENOME Mouse Alox12 siRNA

Protocol tips

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	"SiRNA against 12-LOX was synthesized by a proprietary design as SMARTpool siRNA, which consists of four pooled SMARTselectiondesigned siRNAs (Dharmacon). Cells were electroporated with the siRNAs and plasmids as previously described [17]. Briefly, 5× 105 cells were reconstituted in 100 μl of electroporation transfection solution or Nucleofector Solution V from the Cell Line Nucleofection Kit V (Amaxa Biosystems). siCONTROL or siGenome SMARTpool siRNA against 12- LOX was added to the cells at a final concentration of 200 pmol per sample, and the mixtures were transferred to electroporation cuvettes and subjected to electroporation according to the manufacturer's programs and instructions. The electroporated and transfected cellsuspensions were immediately mixed with 500 μl of prewarmed RPMI medium supplemented with 5% FCS. Cells were then transferred to six-well plates and incubated for 60 h for Western blot analysis or the ESR study"

Protocol tips

"SiRNA against 12-LOX was synthesized by a proprietary design as SMARTpool siRNA, which consists of four pooled SMARTselectiondesigned siRNAs (Dharmacon). Cells were electroporated with the siRNAs and plasmids as previously described [17]. Briefly, 5× 105 cells were reconstituted in 100 μl of electroporation transfection solution or Nucleofector Solution V from the Cell Line Nucleofection Kit V (Amaxa Biosystems). siCONTROL or siGenome SMARTpool siRNA against 12- LOX was added to the cells at a final concentration of 200 pmol per sample, and the mixtures were transferred to electroporation cuvettes and subjected to electroporation according to the manufacturer's programs and instructions. The electroporated and transfected cellsuspensions were immediately mixed with 500 μl of prewarmed RPMI medium supplemented with 5% FCS. Cells were then transferred to six-well plates and incubated for 60 h for Western blot analysis or the ESR study"

Publication protocol

"SiRNA against 12-LOX was synthesized by a proprietary design as SMARTpool siRNA, which consists of four pooled SMARTselectiondesigned siRNAs (Dharmacon). Cells were electroporated with the siRNAs and plasmids as previously described [17]. Briefly, 5× 105 cells were reconstituted in 100 μl of electroporation transfection solution or Nucleofector Solution V from the Cell Line Nucleofection Kit V (Amaxa Biosystems). siCONTROL or siGenome SMARTpool siRNA against 12- LOX was added to the cells at a final concentration of 200 pmol per sample, and the mixtures were transferred to electroporation cuvettes
and subjected to electroporation according to the manufacturer's programs and instructions. The electroporated and transfected cellsuspensions were immediately mixed with 500 μl of prewarmed RPMI medium supplemented with 5% FCS. Cells were then transferred to six-well plates and incubated for 60 h for Western blot analysis or the ESR study"

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