Cignal Reporter Assays include a preformulated, transfection-ready pathway reporter (dual-luciferase or GFP), plus a positive and negative control. The inducible pathway reporter and noninducible negative control are transfected and subjected to experimental treatments in parallel.
Dual-luciferase
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 firefly luciferase and Renilla expression.
GFP
GFP expression is quantified using a flow cytometer, fluorescence microscope, or fluorometer. The change in the activity of each signaling pathway is determined by comparing the GFP activities 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 from the constitutively expressing CMV-GFP reporter. >