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
HEK-293H cells were co-transfected with Oct4-reporter or
negative control (for the transfection protocol, please refer to the user
manual) along with Oct4 expression vector. After 16 hours of transfection medium
was changed to growth medium. After 32 hours of transfection, Dual Luciferase
assay was performed 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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