SwitchGear offers a number of control constructs to optimize experimental conditions and to normalize for non-specific effects associated with a treatment or change in condition. Note that SwitchGear’s unique RenSP luciferase technology eliminates the need for co-transfection of a normalizing control in most cases.
Empty promoter vector: The empty promoter vector contains the RenSP luciferase gene without a promoter. This construct serves as a one measure of background signal in the experiment. This construct may also be used to create additional experimental constructs.
Housekeeping gene promoter vectors: Housekeeping control constructs contain promoters for common housekeeping genes driving the RenSP luciferase reporter. These constructs serve as positive transfection controls and may also serve as controls for comparing signals between conditions if they are known to be unresponsive to the test condition.
Random control vectors: Random control constructs contain 1 kb non-conserved, non-genic, and non-repetitive fragments from the human genome cloned upstream of the luciferase reporter. These vectors produce slightly different signals than the empty vectors and are considered optimal negative or background controls.
Figure 1. Expected data and analysis of housekeeping and random promoter vectors: Random promoter constructs (R1, R2, R3, R4) serve as negative/background controls and establish baseline luciferase activity. The housekeeping promoter controls serve as positive controls and give relative luciferase activity.
Y-axis values shown in log scale. Control vector activity may vary depending on cell line, treatment condition, and protocol changes.