STAT3 Reporter

Research Area

Browse by Pathway

Browse by Disease

STAT3 Reporter

Cignal STAT3 Reporter (luc) Kit: CCS-9028L

The Cignal STAT3 reporter is designed to measure the transcriptional activity of STAT3 homodimers and STA3/STAT1 heterodimers. STAT3 is activated through phosphorylation in response to various cytokines and growth factors including IFNγ, IL-6 and OSM. The activation of STAT3 results in formation of STAT3 homodimers and STAT3/STAT1 heterodimers, which interact with the sis-inducible element (SIE), a promoter sequence, thereby inducing transcription of genes related to cellular processes such as cell growth and apoptosis. The STAT3 reporter is a mixture of a luciferase construct under the control of multiple SIE and a constitutively expressing Renilla construct (40:1). The STAT3-responsive luciferase construct encodes the firefly luciferase reporter gene under the control of a minimal (m)CMV promoter and tandem repeats of the SIE 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. This construct monitors both increases and decreases in the transcriptional activity of STAT3-containing dimers, and hence the activity of the STAT3 signaling pathway. 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, you can easily monitor the activity of the STAT3 signaling pathway and determine the effects of various treatments, such as gene knockdown, over-expression, and chemical compounds on this pathway.

Using a simple dual-luciferase assay, the STAT3 Reporter (luc) can easily and rapidly:

Monitor STAT3 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.

Kit Components

How It Works

Manual & Resources

Related Products

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

HepG2 cells were transfected with STAT3 reporter, negative control and positive control (for transfection protocol, please refer to the user manual). After 16 hours of transfection, medium was changed to growth medium. After 30 hours of transfection, cells were treated with recombinant human Interleukin 6 (hIL-6) for 12 hours. 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