LightSwitch™ Promoter Control Vectors

Synonyms

LightSwitch, Light Switch, reporter assay, reporter gene, luciferase reporter, Renilla luciferase, RenSP, promoter control, random promoter 4, negative control, 32010

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Overview

Because no single promoter can be an ideal control for all experimental conditions, Active Motif offers a panel of positive and negative control constructs for the LightSwitch™ Promoter Reporter Vector. These controls are used to measure background, to optimize experimental conditions and to normalize for non-specific effects associated with a treatment or change in condition. Note that the unique RenSP luciferase technology utilized in the LightSwitch Assay System eliminates the need for co-transfection of a normalizing control in most cases. So, instead of one control construct being co-transfected with every experimental plasmid, different control vectors can be used in your reporter assay experiment, each in its own well, in addition to the constructs being tested. Using the most suitable positive and negative control vectors for your specific experiment will give you the highest degree of confidence in your results.

Information on all LightSwitch Promoter Control Vectors is shown under the Promoter Controls tab below.

IMPORTANT:  Because all LightSwitch reporter constructs contain the optimized RenSP luciferase gene, you MUST use our LightSwitch Luciferase Assay Reagents to obtain optimal results. (Other luciferase assay reagents are not compatible with RenSP.)

Promoter Controls

Empty promoter vector: The empty promoter vector contains the RenSP luciferase reporter gene without a promoter. This construct serves as a measure of background signal in the experiment. This construct may also be used to create additional experimental constructs. For a vector map, annotations and primer & restriction sites, please see the Empty LightSwitch™ Reporter Vectors page.

Housekeeping gene promoter vectors: Housekeeping control constructs contain promoters for common housekeeping genes driving the RenSP luciferase reporter gene. 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 promoter vectors: Random control constructs contain 1 kb non-conserved, non-genic, non-repetitive fragments from the human genome cloned upstream of the RenSP luciferase reporter gene. These vectors produce slightly different signals than the empty promoter vector and are considered optimal negative or background controls.

Figure 1: LightSwitch assay performed using the positive and negative LightSwitch Promoter Controls.The 4 housekeeping promoter controls serve as positive controls and give relative luciferase activity. The 4 random promoter constructs and the Empty LightSwitch Reporter Vector serve as negative/background controls and establish baseline luciferase activity.
The Y-axis values are shown in log scale. Control vector activity may vary depending on cell line, treatment condition and protocol changes.

Promoters of housekeeping genes (Positive controls)

Catalog No.	Name	Description	Coordinates	Sequence
32003	LightSwitch™ ACTB Promoter Control	Beta-actin	chr7–: 5343428 - 5344428	Get Info
32004	LightSwitch™ GAPDH Promoter Control	Glyceraldehyde-3-phosphate dehydrogenase	chr12+: 6513163 - 6514226	Get Info
32005	LightSwitch™ LDHA Promoter Control	Lactose dehydrogenase A	chr11+: 18372059 - 18372891	Get Info
32006	LightSwitch™ RPL10 Promoter Control	Ribosomal protein L10	chrX+: 153146645 - 153147658	Get Info

Random genomic fragments in promoter vector (Negative controls)

Catalog No.	Name	Coordinates	Sequence
32007	LightSwitch™ Random Promoter Control 1	chr11+: 119015626 - 119016592	Get Info
32008	LightSwitch™ Random Promoter Control 2	chr1+: 226079717 - 226080649	Get Info
32009	LightSwitch™ Random Promoter Control 3	chr6+: 15604416 - 15605350	Get Info
32010	LightSwitch™ Random Promoter Control 4	chr16+: 1645930 - 1646850	Get Info

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