Oct4 Reporter

Cignal Oct4 Reporter (luc) Kit: CCS-0025L The Cignal Oct4 reporter is designed to measure the transcriptional activity of Oct4. Oct-4 is a transcription factor whose expression level governs the fate of the cells within the primitive inner cell mass and embryonic stem (ES) cells. The Oct4 reporter is a mixture of an Oct4-responsive luciferase construct and a constitutively expressing Renilla construct (40:1). The Oct4-responsive luciferase construct encodes the firefly luciferase reporter gene under the control of a minimal (m)CMV promoter and tandem repeats of the Oct4 transcriptional response element. The number of response elements and the intervening sequence between these response elements has been experimentally optimized to maximize the signal to noise ratio. The Oct4 reporter construct monitors both increases and decreases in the transcriptional activity of Oct4. The constitutively expressing Renilla construct encodes the Renilla luciferase reporter gene under the control of a CMV immediately early enhancer/promoter and acts as an internal control for normalizing transfection efficiencies and monitoring cell viability. Using a simple dual-luciferase assay, the activity of Oct4 can be easily measured in response to various treatments, such as gene knockdown, over-expression, and chemical compounds. Using a simple dual-luciferase assay, the Oct4 Reporter (luc) can easily and rapidly: Monitor Oct4 transcriptional activity in cells Study chemical compounds or drugs Analyze phenotypes of RNAi or gene over expression See the complete transcription factor reporter assay list.

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10-Pathway Arrays Component Specification Concentration* (total volume) Oct4 Reporter  A mixture of inducible Oct4-responsive firefly luciferase construct and constitutively expressing Renilla luciferase construct (40:1).  (100 ng/µl; 500 µl) Negative control  A mixture of non-inducible firefly luciferase construct and constitutively expressing Renilla luciferase construct (40:1).  (100 ng/µl; 500 µl) Positive control  A mixture of constitutively expressing GFP, constitutively expressing firefly luciferase, and constitutively expressing Renilla luciferase constructs (40:1:1).  (100 ng/µl; 250 µl)

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Cignal Technology Overview Brief Protocol The Cignal Reporter Assays (luc) include pre-formulated, transfection-ready reporter, negative control, and positive control. The transcription factor reporter and negative control are transfected and subjected to experimental treatments, in parallel. 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 transfectants serve 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. Performance Data Excellent signal to noise ratio HEK-293H cells were co-transfected with Oct4-reporter or negative control (for the transfection protocol, please refer to the user manual) along with Oct4 expression vector. After 16 hours of transfection medium was changed to growth medium. After 32 hours of transfection, Dual Luciferase assay was performed and promoter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were done in triplicates, and the standard deviation is indicated. Back to Top

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User Manuals Cignal Reporter Assay Kit User Manual (PDF) White Paper Cignal Reporter Assay Kit: A High Performance Tool for Assessing the Functions of Genes, Biologics and Small Molecule Compounds (PDF) Web Seminar Attend a live on-line seminar hosted by an Application Scientist about Cignal Reporters Technical Brochure Cignal Reporter Assay Kits Technical Brochure (PDF) Power Point Cignal™ Cell-Based Assays for Rapidly Analyzing Signaling Pathway Activity (PDF) Back to Top

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Pathway reporters Complete list of signaling pathway reporters Attractene Transfection Achieve high transfection efficiencies and minimal cytotoxicity in your transfections 10-Pathway Arrays Simultaneously measure the activities of ten (10) signaling pathways in a single experiment shRNA Plasmids Pathway related gene knockdowns - Guaranteed 70% gene knockdown Back to Top