Would you like to pinpoint the signaling pathways regulated by your gene product or compound? If so, consider Cignal Finder Reporter Arrays - the first commercial products that enable a comprehensive analysis of multiple pathways (up-to ten) in a single experiment.

• Learn how the reporter arrays work
  In a single experiment you can use the Cignal Finder 10-Pathway Reporter Arrays to simultaneously measure the activities of 10 cell signaling pathways known to play critical roles in the biological process being studied in your laboratory. Don't rely on guesswork to predict mechanisms of action. Use the Cignal Finder Arrays to definitively discover important gene functions and pathway interactions.
  
  
• See an application experiment

Which ten pathways? See the Cignal Finder Array pathway lists

The Cignal Finder 10-Pathway Reporter Arrays are ideally suited for discovering:

• siRNA/shRNA/miRNA phenotypes
• Biological responses to chemical compounds
• Mechanisms of action of proteins, peptides, and ligands

Available Cignal Finder 10-Pathway Reporter Arrays (luciferase)

Why Cignal Finder 10-Pathway Reporter Arrays?

• Biological Process-Focused: Profile changes in the activities of ten signaling pathways relevant to a specific biological process
• High Performance: Dual-luciferase assay method provides high sensitivity, specificity, and reproducibility
• Flexibility and Convenience: Utilize a straightforward traditional transfection or reverse transfection procedure with your favorite cell lines to rapidly generate valuable mechanism of action data

How It Works

Each Reporter Array includes 10 Cignal Reporter Assays and two controls in either 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.

	Simple Procedure: Transfect Cignal Reporter Assays and test nucleic acids into cells Treat with protein, peptide, or small molecule of interest Perform reporter quantitation using luciferase activity assays
Cignal Finder Array - Tube Format The tube format of the Cignal Finder Arrays are delivered in 12-tube strips, along with important negative and positive controls. The assays are used right out of the box for the transfection or reverse transfection of the reporter assays into your cell lines of interest.
Cignal Finder Array - Plate Format The plate format of the Cignal Finder Arrays 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).

Application Example

Identification of signaling pathway(s) impacted by p53 siRNA treatment
The most heavily studied tumor suppressor gene is p53. To understand more about the biological function of p53 in the cell line of your choice, it is important to know the signaling pathways perturbed by knockdown of p53. The Cignal Finder Cancer 10-pathway Reporter Array provides a vital tool to identify the key cancer signaling pathways modulated by knock down of p53.

The Cignal Finder Cancer 10-Pathway Reporter Array showed that the knock down of p53 gene expression down-regulates p53 signaling, while up-regulating Notch, hypoxia and MAPK/ERK signaling in HEK-293H cells. Interestingly, Notch signaling is known to be frequently deregulated in human malignancies. Up-regulation of Notch signaling by p53 RNA interference suggests that Notch may function as a proto-oncogene.

HEK-293H cells were co-transfected 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. Sixteen hours after carrying out the reverse transfection, medium was changed to complete medium (DMEM containing 10% of fetal bovine serum, 1% NEAA, 100 U/ml Penicillin and 100 µg/ml Streptomycin). After 48 hours of transfection, the dual-luciferase assay was performed and results are expressed as fold change. The fold change was calculated by dividing the normalized luciferase activities of each pathway-focused reporter co-transfected with p53 siRNA by the normalized luciferase activity of each pathway-focused reporter co-transfected with the negative control siRNA. Experiments were done in quadruplicates, and the standard deviation is indicated.