Cignal Reporter Controls

For control experiments when using Cignal Reporter Assays

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Cignal Reporter Controls

Cat. No. / ID: 336881

Cignal Reporter Control

Cignal Reporter Controls are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

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Find or custom design the right target-specific assays and panels to research your biological targets of interest.

Features

Highly suited for optimizing transfection conditions
Establish the specificity of any treatment effects
Transfection-ready constructs
High sensitivity and specificity

Product Details

Cignal Reporter Assays provide a rapid, sensitive, and quantitative assessment of signal transduction pathway activation by measuring the activities of downstream transcription factors. When conducting experiments using Cignal Reporter Assays, proper controls are a key element of the experimental design. They permit accurate interpretation of reporter assay results and provide assurance of the specificity of the observed response.

Principle

Cignal Reporter Controls are preformulated, transfection-ready constructs. Positive and negative controls are available for both dual-luciferase and GFP reporters.

The following controls are available.

Available controls
Control	Description
Positive Control Assay (GFP)	Constitutively expressing GFP gene. Easily measure transduction efficiency and optimize transduction conditions with green fluorescent protein (GFP)
Negative Control Assay (GFP)	Noninducible, minimal promoter-driven GFP gene. Establish the specificity of any treatment effects and determine background GFP fluorescence
Positive Control Assay (luc)	40:1:1 mixture of constitutively expressing GFP, constitutively expressing firefly luciferase, and constitutively expressing Renilla luciferase constructs. Measure transduction efficiency and serve as positive control for firefly luciferase assay
Negative Control Assay (luc)	40:1 mixture of non-inducible firefly luciferase construct and constitutively expressing Renilla luciferase construct. Establish the specificity of any treatment effects

Procedure

Cignal Lenti Reporter Control Assays are pretitered, transduction-ready lentivirus particles used for optimizing transduction conditions. Use standard procedures to transduce Cignal Lenti Reporter Control Assays into selected cells, and treat control-transduced cells in the same way as reporter-transduced cells.

Applications

Cignal Reporter Controls are a key element of the experimental design when conducting experiments using Cignal Reporter Assays. They are versatile tools for:

Optimizing transfection conditions for a cell system
Quantifying transfection efficiency
Establishing the specificity of any treatment effects
Determining background GFP fluorescence or luciferase activity
Serving as an internal control for normalization

Supporting data and figures

Cignal Reporter Controls.

Resources

Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.

Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.

Download Safety Data Sheets for QIAGEN product components.

For cell-based pathway activity assays

For cell-based analysis of pathway signaling activity

FAQ

Any transfection reagent that yields 25-30% transfection efficiency in the cell line of interest is suitable for use with DNA-based Cignal Reporter Assays.

FAQ ID -2764

No, The Cignal Reporter Assays do not currently support this functionality.

FAQ ID -2765

No, DNA-based Cignal Reporter Assays are not designed for bacterial transformations.

FAQ ID -2762

No, the positive control only serves as a constitutively-active reporter enabling the user to assess if the transfection/transduction was successful, and should not be used for direct comparison to the pathway reporters.

FAQ ID -2766

MOI is an abbreviation for Multiplicity of Infection or the number of viral particles exposed to a cell.

FAQ ID -2768

This has to be empirically determined by the user. By using a positive control, Cignal Lenti Reporter Control, the user can set up parallel transductions with increasing amounts of virus to identify an MOI that yields a robust signal.

FAQ ID -2770

No, The negative control only serves as a non-inducible reporter, but should not be used for direct comparison to the pathway reporters.

FAQ ID -2767

No, the DNA-based Cignal Reporter Assays are not designed to generate stable pathway sensor cell lines. However, the Cignal Lenti Reporter Assays can be used to generate stable pathway sensor cell lines.

FAQ ID -2763

The concentration of SureENTRY Transduction Reagent is 10 mg/ml.

FAQ ID - 3708

The DNA-based Reporters are designed for cells that are amenable to transfection. QIAGEN recommends a minimum transfection efficiency of 25 – 30%. If the cells are refractory to transfection then the Cignal Lenti reporters should be selected.

FAQ ID -2761

This depends on the Multiplicity of Infection (MOI) required for a specific cell line. The lower the MOI the more cells that can be transduced. Therefore, to predict how much Cignal Lenti Reporter is required an MOI must be determined.

FAQ ID -2769