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We currently have validated protocols (≥70% mRNA knockdown of GAPDH with ≥70% relative cell viability) for the following cell types:
3T3-L1 Differentiated mouse adipocytesshow
Differentiated mouse adipocytes were plated at 10,000 cells/well in a 96-well plate
overnight and then transfected with varying concentrations (1.9-30 nM) of Mouse
GAPDH siRNA Positive Control (Ambion, P/N 4624). Twenty-four hours post transfection,
cells were lysed and GAPDH and cyclophillin B (PPIB) gene expression was quantitated
using the QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
C2C12 Differentiated mouse myotubesshow
Mouse C2C12 cells were plated at 3,000 cells/well in a 96-well plate, differentiated into myotubes for 4 days, and then transfected with varying concentrations (12.5-100 nM) of Mouse GAPDH siRNA Positive Control (Ambion, P/N 4624). Twenty-four hours post
transfection, cells were lysed and GAPDH and cyclophillin B (PPIB) gene expression was
quantitated using the QuantiGene Reagent System. Relative cell viability was >70% for 12.5-75 nM siRNA concentrations evaluated.
C2C12 Undifferentiated mouse myocytesshow
Undifferentiated C2C12 cells were plated at 3,000 cells/well in a 96-well plate overnight and then transfected with varying concentrations (0.8-30 nM) of Mouse GAPDH siRNA Positive Control (Ambion, P/N 4624). Twenty-four hours post transfection, cells were lysed and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
RAW 264.7 Mouse macrophage cellsshow
RAW 264.7 cells were plated at 8,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Mouse GAPDH siRNA Positive
Control (Ambion, P/N 4624). Twenty-four hours post transfection, cells were lysed and
GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the QuantiGene
Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
U87MG Human brain glioblastoma astrocytoma cellsshow
U87MG cells were plated at 10,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
Astrocytoma cells, humanshow
Astrocytoma cells were plated at 10,000 cells/well in a 96-well plate overnight and then transfected with varying concentrations (2.5-10 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
NHEK-AD Primary human adult keratinocytesshow
NHEK-AD cells were plated at 150,000 and 300,000 cells/well in a 6-well plate overnight and then transfected with 20 nM of Human GAPDH siRNA Positive Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the QuantiGene Reagent
System. Relative cell viability was >70% for the 20 nM siRNA concentration evaluated at
both cell densities.
THP-1 Human peripheral blood acute monocytic leukemia cellsshow
THP-1 cells were plated at 10,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.625-20 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for 0.625-10 nM siRNA concentrations evaluated.
HUVEC Human umbilical vein endothelial cellsshow
HUVEC were plated at 10,000 cells/well in a 96-well plate overnight and then transfected
with 15 nM of Human GAPDH siRNA Positive Control (Panomics P/N DX0101). The
incubation time with the transfection complex, serum-free media and cells was either 30
minutes or 2 hours. Twenty-four hours post transfection, cells were lysed and GAPDH
and cyclophillin B (PPIB) gene expression was quantitated using the QuantiGene Reagent
System. Relative cell viability was >70% at the 15 nM siRNA concentration evaluated.
MDA-MB-231 Human breast adenocarcinoma cellsshow
31 cells were plated at 10,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for 0.8-15 nM siRNA concentrations evaluated.
MCF-7 Human breast carcinoma cellsshow
MCF-7 cells were plated at 5,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
HT29 Human colon carcinoma cellsshow
HT29 were plated at 5,000 cells/well in a 96-well plate overnight and then transfected
with varying concentrations (0.1-20 nM) of Human GAPDH siRNA Positive Control
(Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed and
GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the QuantiGene
Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
SW620 Human colon carcinoma cellsshow
SW620 cells were plated at 5,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.1-20 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for 0.1-10 nM siRNA concentrations evaluated.
HepG2 Human hepatocarcinoma cellsshow
HepG2 cells were plated at 10,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (1.25-20 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for 1.25-5 nM siRNA concentrations evaluated.
BSMC Human bronchial smooth muscle cellsshow
BSMC cells were plated at 6,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (5-20 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
A549 Human lung carcinoma cellsshow
A549 cells were plated at 10,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
A2780 Human ovarian cancer cellsshow
A2780 cells were plated at 5,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.8-30 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for 0.8-15 nM siRNA concentrations evaluated.
ASPC-1 Human pancreatic carcinoma cellsshow
ASPC-1 cells were plated at 5,000 cells/well in a 96-well plate overnight and then
transfected with varying concentrations (0.625-20 nM) of Human GAPDH siRNA Positive
Control (Panomics, P/N DX0101). Twenty-four hours post transfection, cells were lysed
and GAPDH and cyclophillin B (PPIB) gene expression was quantitated using the
QuantiGene Reagent System. Relative cell viability was >70% for all siRNA concentrations evaluated.
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