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The Cignal Reporter Assays (luc) include pre-formulated, transfection-ready reporter, negative control, and positive control. The transcription factor reporter and negative control are transfected and subjected to experimental treatments, in parallel. Dual-luciferase results are calculated for each transfectant. The change in the activity of each signaling pathway is determined by comparing the normalized luciferase activities of the reporter in treated versus untreated transfectants. The identically treated negative control transfectants serve as a specificity control. The positive control serves as a control for transfection efficiency, by monitoring GFP expression, as well as a positive control for both the firefly and Renilla luciferase assays.
Excellent signal to noise ratio
HepG2 cells were co-transfected with PPAR-reporter or
negative control (for transfection protocol, please refer to the user
manual) along with PPAR gamma expression vector. After 16 hours of
transfection medium was changed to growth medium. After 24 hours of
transfection, cells were treated with 1 µM of rosaglitazone. Dual
Luciferase assay was performed 18 hours after treatment, and promoter
activity values are expressed as arbitrary units using a Renilla reporter
for internal normalization. Experiments were done in triplicates, and the
standard deviation is indicated.
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