Androgen Receptor Reporter

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Cignal Androgen Receptor Reporter (luc) Kit: CCS-1019L

The AR reporter is designed to measure the transcriptional activity of the androgen receptor (AR). The androgen receptor, also known as NR3C4 (nuclear receptor subfamily 3, group C, member 4), is a nuclear receptor that is activated by binding androgenic hormones, testosterone or dihydrotestosterone. In its ligand bound state, AR dimerizes and regulates gene expression by binding to specific hormone response elements. The AR reporter is a mixture of an androgen-responsive luciferase construct and a constitutively expressing Renilla construct (40:1). The AR-responsive luciferase construct encodes the firefly luciferase reporter gene under the control of a minimal (m)CMV promoter and tandem repeats of the AR 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 AR reporter construct monitors both increases and decreases in the transcriptional activity of the androgen receptor, and hence the activity of the androgen signaling. 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 androgen receptor and determine the effect of various treatments, such as gene knockdown, over-expression, and chemical compounds.

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

Monitor Androgen Receptor 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

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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

LNCaP cells were transfected with AR reporter (200 ng), negative control and positive control (for the transfection protocol, please refer to the user manual). After 16 hours of transfection, medium was changed to assay medium (Opti-MEM + 1% FBS + 0.1mM NEAA). After 24 hours of transfection, cells were treated with Mibolerone, a synthetic androgen, (3.2 nM) for 18 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.

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