ON-TARGETplus Human BAK1 (578) siRNA - SMARTpool

Protocol tips

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	Cells were plated in 6-well plates at 2.0 ×105 cells per well in RPMI complete media. The following day the media was replaced with 1ml Opti-MEM (Invitrogen) and the cells were transfected with 0.5ml complexes of Lipofectamine 2000 (0.5%, Invitrogen) and siRNA for either Mcl-1, BIM, BAK, Wee1 or non-targeting controls (40nM, Dharmacon On-Target Plus pools). Medium was replaced after 3–6 hours with 5% FBS in RPMI, and the cells were analyzed 120 hr after transfection. In the BIM and BAK studies, 300nM XL888 was also added 48 hours after transfection.

Protocol tips

Cells were plated in 6-well plates at 2.0 ×105 cells per well in RPMI complete media. The following day the media was replaced with 1ml Opti-MEM (Invitrogen) and the cells were transfected with 0.5ml complexes of Lipofectamine 2000 (0.5%, Invitrogen) and siRNA for either Mcl-1, BIM, BAK, Wee1 or non-targeting controls (40nM, Dharmacon On-Target Plus pools). Medium was replaced after 3–6 hours with 5% FBS in RPMI, and the cells were analyzed 120 hr after transfection. In the BIM and BAK studies, 300nM XL888 was also added 48 hours after transfection.

Publication protocol

Cells were plated in 6-well plates at 2.0 ×105 cells per well in RPMI complete media. The following day the media was replaced with 1ml Opti-MEM (Invitrogen) and the cells were transfected with 0.5ml complexes of Lipofectamine 2000 (0.5%, Invitrogen) and siRNA for either Mcl-1, BIM, BAK, Wee1 or non-targeting controls (40nM, Dharmacon On-Target Plus pools). Medium was replaced after 3–6 hours with 5% FBS in RPMI, and the cells were analyzed 120 hr after transfection. In the BIM and BAK studies, 300nM XL888 was also added 48 hours after transfection.

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