Cignal 45-Pathway Reporter Array
For discovery of signaling pathway response to gene modification or chemical treatment
The Cignal 45-Pathway Reporter Array measures the activity of 45 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. The plate format facilitates shorter transfection protocols and high-throughput analysis.
Principle
Each array includes 45 Cignal Reporter Assays and 2 controls in a 96-well plate format. Each reporter and control assay is dried down in each well of the plate.
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.
Procedure
The procedure comprises 3 simple steps (see figure "Cignal 45-Pathway Reporter Array procedure"):
Applications
The Cignal 45-Pathway Reporter Array is 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. |
