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Cignal Finder Reporter Arrays

For discovery of signaling pathway response to gene modification or chemical treatment

  • Analysis of multiple pathways in a single experiment
  • Flexible formats
  • Assays gene knockdown and overexpression
  • Assays chemical modulators

Cignal Finder Reporter Array allows for the comprehensive analysis of 10 signaling pathways. By screening pathway activities simultaneously, relevant pathways are quickly identified for further analysis. Cignal Finder Reporter Arrays pinpoint the pathways perturbed by a specific gene or drug.

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

Signaling pathways affected by p53 knockdown.

HEK-293H cells were cotransfected with either p53 siRNA or a negative control siRNA, in combination with each reporter assay and the negative control from the Cancer 10-Pathway Reporter Array plate. 16 hours after transfection, medium was changed to complete medium. 48 hours after transfection, the dual-luciferase assay was performed. Results are expressed as fold change.

Signaling pathways affected by TNFα.

HeLa cells were reverse transfected with the Immune Response 10-Pathway Cignal Finder Reporter Array. 16 hours after transfection, medium was changed to assay medium. 32 hours after transfection, cells were treated with 5 ng/ml TNFα or left untreated. After 6 hours treatment, dual-luciferase assays were performed. Results are expressed as fold change.

Cignal Finder Array, tube format procedure.
Cignal Finder Array, plate format procedure.
Identification of optimal drug dose.

HepG2 cells were reverse transfected with each reporter assay and the controls on the Stress and Toxicity 10-Pathway Reporter Array plate. 16 hours after transfection, the medium was changed to complete medium. After 30 hours, cells were treated with increasing dosages of tunicamycin (0–4 µg/ml). After 48 hours, the dual-luciferase assay was performed; results are expressed as fold change. 0.125 µg/ml Tunicamycin is the lowest effective non-toxic dose for glycoprotein synthesis inhibition.


Cignal Finder Reporter Arrays can be used in many applications.

Identification of optimal drug doses

Toxicity is one of the primary reasons potential drug candidates fail during development. Identifying potential toxicity and the cellular response to a given concentration or compound can benefit future studies. Toxic compounds or dosages activate various stress response pathways. Cell-based luciferase reporter assays were used to measure 10 stress response pathways using a single 96-well plate.

Tunicamycin inhibits the enzyme GlcNAc phosphotransferase (GPT) and induces endoplasmic reticulum (ER) stress. In this study, the Stress and Toxicity 10-Pathway Reporter Array was used to determine the lowest effective concentration of Tunicamycin without activating cellular stress pathways. Results showed that 0.125 µg/ml Tunicamycin is the lowest effective non-toxic dose for glycoprotein synthesis inhibition (see figure "Identification of optimal drug dose").

Cell response to loss of a cancer signaling pathway

The most well-studied tumor suppressor gene is p53. To understand more about the biological function of p53, it is important to measure the signaling pathways affected by p53 knockdown. The Cancer 10-Pathway Reporter Array can help to identify key cancer signaling pathways modulated by p53 knockdown (see figure "Signaling pathways affected by p53 knockdown").

Results showed that knockdown of p53 gene expression downregulates p53 signaling, while upregulating Notch, hypoxia, and MAPK/ERK signaling in HEK-293H cells. Notch signaling is known to be frequently deregulated in human malignancies. Activation of Notch signaling by p53 RNA interference suggests that Notch may function as a proto-oncogene.

Measurement of signaling in response to cytokines

Tumor necrosis factor alfa (TNFα) is a pleiotropic inflammatory cytokine. It is important to determine the key immunology signaling pathways modulated by TNFα. The Immune Response 10-Pathway Cignal Finder Reporter Array may provide valuable information about the signaling pathways involved in the biological response to TNFα.

The Immune Response 10-Pathway Cignal Finder Reporter Array reveals that TNFα activates the NFkB and MAPK/JNK signaling pathways in HeLa cells (see figure Signaling pathways affected by TNFalpha).


Each array includes 10 Cignal Reporter Assays and 2 controls in either a transfection-ready tube or plate format.
All reporter assays are based on dual-luciferase technology. Each reporter consists of a mixture of a pathway-focused transcription factor-responsive firefly luciferase construct and a constitutively expressing Renilla luciferase construct.

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 serves 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.


The procedure is comprised of 3 simple steps:

Transfect Cignal Finder Reporter Assays and test nucleic acids into cells 
Treat with siRNA, protein, peptide, or small molecule of interest 
Perform reporter quantitation using luciferase activity assays
Cignal Finder Reporter Array: Tube Format

Cignal Finder Reporter Arrays in tube format are delivered in 12-tube strips, along with positive and negative controls. The assays are immediately ready for use in the transfection or reverse transfection of the reporter assays into the cell lines of interest (see flowchart "Cignal Finder Array, tube format procedure").

Cignal Finder Reporter Array: Plate Format

Cignal Finder Reporter Arrays in plate format are delivered in a 96-well cell culture plate. Each reporter and control assay is dried down in each column of the plate (8 wells per assay; see flowchart "Cignal Finder Array, plate format procedure").


Cignal Reporter Arrays are highly suited for research into phenotypes associated with RNAi or overexpression experiments, biological responses to small molecules or compounds, and mechanisms of action of proteins, peptides, and ligands.

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Brochures & Guides (1)
For cell-based analysis of pathway signaling activity
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Kit Handbooks (3)
Transfection Protocols (2)
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.
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The Cignal Signal Transduction 45-Pathway Reporter Array enables comprehensive analysis of 45 signaling pathways, including pathways involved in immune signaling, hormone signaling, and more. Screening a large number of pathways simultaneously helps pinpoint the pathways perturbed by a specific gene or drug, quickly identifying relevant pathways for further analysis.
Plate layout
Well position Pathway Transcription factor Well position Pathway Transcription factor
A1–2 Amino Acid Deprivation ATF2/3/4 E1–2 MAPK/Jnk AP-1
A3–4 Androgen AR E3–4 MEF2 MEF2
A5–6 Antioxidant Response Nrf2/Nrf1 E5–6 Myc c-Myc
A7–8 ATF6 ATF6 E7–8 Nanog Nanog
A9–10 C/EBP C/EBP E9–10 Notch RBP-Jk
A11–12 cAMP/PKA CREB E11–12 NFκB NFκB
B1–2 Cell Cycle E2F F1–2 Oct4 Oct4
B3–4 DNA Damage p53 F3–4 Pax6 Pax6
B5–6 EGR1 EGR1 F5–6 PI3K/Akt FOXO
B7–8 ER Stress CBF/NF-Y/YY1 F7–8 PKC/Ca++ NFAT
B9–10 Estrogen ER F9–10 PPAR PPAR
B11–12 GATA GATA F11–12 Progesterone PR
C1–2 Glucocorticoid GR G1–2 Retinoic Acid RAR
C3–4 Heat Shock HSF-1 G3–4 Retinoid X RXR
C5–6 Heavy Metal MTF-1 G5–6 Sox2 Sox2
C7–8 Hedgehog Gli G7–8 SP1 SP1
C9–10 HNF4 HNF4 G9–10 STAT3 STAT3
C11–12 Hypoxia HIF-1a G11–12 TGF-β SMAD2/3/4
D1–2 Interferon Regulation IRF1 H1–2 Vitamin D VDR
D3–4 Type I Interferon STAT1/STAT2 H3–4 Wnt TCF/LEF
D5–6 Interferon-γ STAT1 H5–6 Xenobiotic AhR
D7–8 KLF4 KLF4 H7–9 Negative Control  
D9–10 Liver X LXR H10–12 Positive Control  
D11–12 MAPK/Erk SRF/Elk-1      

Name Cignal Finder Signal Transduction 45-pathway Reporter Array (CCA-901L)