A total of 1.2 × 104 cells were plated into 12-well cell culture plates and incubated at 37 °C and 5% CO2 overnight. The following day, cells were transfected with 5 µl of 5 µM siRNA with 5 µl of DharmaFECT 1 Transfection Reagent (Horizon Discovery). Cells were either transfected with ON-TARGETplus MDH1 siRNA (L-051206-01-0005; Horizon Discovery) or ON-TARGETplus non-targeting control siRNA (D-001810-10-05; Horizon Discovery). Cells were supplemented with excess media the following day and metabolites were extracted 48 h post transfection as outlined above. Mass spectrometry and subsequent targeted metabolomics analysis was performed as previously described38 (link), using Tracefinder (v.5.1). Compound peak areas were normalized using the total measured protein per well quantified with a modified Lowry assay38 (link).
On targetplus mdh1 sirna
Manufactured by Horizon Discovery
The ON-TARGETplus MDH1 siRNA is a laboratory reagent designed for gene silencing experiments. It is a double-stranded RNA molecule that targets the MDH1 gene, which encodes the malate dehydrogenase 1 enzyme. The siRNA can be used to temporarily reduce the expression of the MDH1 gene in cell cultures or other experimental systems.
Automatically generated - may contain errors
2 protocols using on targetplus mdh1 sirna
1
Targeted Metabolomic Analysis of siRNA Knockdown
2
Metabolic impact of MDH1 knockdown
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1.2 × 124 cells were plated into 12-well cell culture plates and incubated at 37°C and 5% CO2 overnight. The following day, cells were transfected with 5µL of 5µM siRNA with 5µL of DharmaFECT 1 Transfection Reagent (Horizon Discovery). Cells were either transfected with ON-TARGETplus MDH1 siRNA (L-051206–01-0005, Horizon Discovery) or ON-TARGETplus non-targeting control siRNA (D-001810–10-05, Horizon Discovery). Cells were supplemented with excess media the following day and metabolites extracted 48hrs post-transfection as outlined above.
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