siGENOME Rat Sod2 (24787) siRNA - SMARTpool

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	Normal rat kidney (NRK, ATCC CRL 6509) cells were maintained in a humidified incubator with 5% CO2, 95% air at 37 °C in DMEM containing 5% fetal calf serum, and grown to 70% confluency. MnSOD siRNA (siGENOME siRNA SMARTpool, Dharmacon) was used to knockdown MnSOD in the cells. A nonsense siRNA (siGENOME NON-TARGETing siRNA #2, Dharmacon) was used as a negative control. Briefly, the siRNA was diluted (5–25 nM final concentration) in OptiMem and incubated (25 min, 25 °C) with lipofectamine (Invitrogen) prior to adding to cells containing DMEM only. Cells were incubated with the siRNA solution for 6 h and then placed back in normal media. Successful knockdown of MnSOD was confirmed by measuring MnSOD expression and activity compared to cells treated with nonsense siRNA. The percentage of transfection efficiency was 98% using BLOCK-iT fluorescent Oligo uptake at 24 h post transfection (data not shown).

Protocol tips

Normal rat kidney (NRK, ATCC CRL 6509) cells were maintained in a humidified incubator with 5% CO2, 95% air at 37 °C in DMEM containing 5% fetal calf serum, and grown to 70% confluency. MnSOD siRNA (siGENOME siRNA SMARTpool, Dharmacon) was used to knockdown MnSOD in the cells. A nonsense siRNA (siGENOME NON-TARGETing siRNA #2, Dharmacon) was used as a negative control. Briefly, the siRNA was diluted (5–25 nM final concentration) in OptiMem and incubated (25 min, 25 °C) with lipofectamine (Invitrogen) prior to adding to cells containing DMEM only. Cells were incubated with the siRNA solution for 6 h and then placed back in normal media. Successful knockdown of MnSOD was confirmed by measuring MnSOD expression and activity compared to cells treated with nonsense siRNA. The percentage of transfection efficiency was 98% using BLOCK-iT fluorescent Oligo uptake at 24 h post transfection (data not shown).

Publication protocol

Normal rat kidney (NRK, ATCC CRL 6509) cells were maintained in a humidified incubator with 5% CO2, 95% air at 37 °C in DMEM containing 5% fetal calf serum, and grown to 70% confluency. MnSOD siRNA (siGENOME siRNA SMARTpool, Dharmacon) was used to knockdown MnSOD in the cells. A nonsense siRNA (siGENOME NON-TARGETing siRNA #2, Dharmacon) was used as a negative control. Briefly, the siRNA was diluted (5–25 nM final concentration) in OptiMem and incubated (25 min, 25 °C) with lipofectamine (Invitrogen) prior to adding to cells containing DMEM only. Cells were incubated with the siRNA solution for 6 h and then placed back in normal media. Successful knockdown of MnSOD was confirmed by measuring MnSOD expression and activity compared to cells treated with nonsense siRNA. The percentage of transfection efficiency was 98% using BLOCK-iT fluorescent Oligo uptake at 24 h post transfection (data not shown).

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