FACS sorted fibroblasts were cultured in DMEM supplemented with 10% FCS. At 70% confluency, cells were transfected with Accell Delivery Media (GE Dharmacon; B-005000) supplemented with 1 µM Accell SMARTpool mouse Clusterin siRNA (GE Dharmacon; E-043966-00) or Accell Control Pool nontargeting siRNA (GE Dharmacon; D-001910-10) for 48 h. Accell SMARTpool contains a mixture of four siRNAs targeting one gene and provides extended duration of gene knockdown with only minimal effects on cell viability and the innate immune response. The efficiency of Clusterin siRNA knockdown was analyzed by qRT-PCR.
Accell control pool nontargeting sirna
Manufactured by Horizon Discovery
Accell Control Pool nontargeting siRNA is a pool of four unique siRNA duplexes designed as a negative control for siRNA experiments. The pool does not target any known gene and can be used to monitor nonspecific effects in cell-based assays.
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Lab products found in correlation
2 protocols using accell control pool nontargeting sirna
1
Fibroblast Clusterin Knockdown Protocol
2
Myc Knockdown and Overexpression in NLFs
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NLFs were cultured in DMEM supplemented with 10% FCS. At 70% confluency, cells were transfected with Accell Delivery Media (GE Dharmacon; B-005000) supplemented with 1 µM Accell SMARTpool mouse Myc siRNA (Dharmacon; E-040813) or Accell Control Pool nontargeting siRNA (Dharmacon; D-001910) for 96 hr. Accell SMARTpool contains a mixture of four siRNAs targeting one gene and provides extended duration of gene knockdown with only minimal effects on cell viability and the innate immune response. The efficiency of Myc siRNA knockdown was analyzed by qRT-PCR.
For individual siRNA experiments, NLFs were cultured and transfected as described, utilizing individual Myc targeting siRNA constructs (Dharmacon, A-040813-20, A-040813-18, A-040813-17) or control siRNA.
For overexpression of Myc, cells were transiently transfected with a plasmid of Myc (MGC Mouse Myc cDNA pCMV-SPORT6: mammalian expression insert sequence: BC006728, #MMM1013-202763479) or with a control plasmid. Cells were transfected with jetPRIME (polyplus transfection, 114-01) according to the manufacturer’s protocol. All experiments were performed 24 hr following transfection.
XTT assay (Biological Industries, 20-300-1000) was performed 24 hr following transfection according to the manufacturer’s protocol.
For individual siRNA experiments, NLFs were cultured and transfected as described, utilizing individual Myc targeting siRNA constructs (Dharmacon, A-040813-20, A-040813-18, A-040813-17) or control siRNA.
For overexpression of Myc, cells were transiently transfected with a plasmid of Myc (MGC Mouse Myc cDNA pCMV-SPORT6: mammalian expression insert sequence: BC006728, #MMM1013-202763479) or with a control plasmid. Cells were transfected with jetPRIME (polyplus transfection, 114-01) according to the manufacturer’s protocol. All experiments were performed 24 hr following transfection.
XTT assay (Biological Industries, 20-300-1000) was performed 24 hr following transfection according to the manufacturer’s protocol.
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