EGR1 Reporter

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EGR1 Reporter

Cignal EGR1 Reporter (luc) Kit: CCS-8021L

The EGR1 reporter is designed to measure the transcriptional activity of the early growth response 1 (EGR1) transcription factor, which belongs to the EGR family of C2H2-type zinc-finger proteins. The target genes of EGR1, a nuclear protein, are required for differentitation and mitogenesis. The EGR1 reporter is a mixture of an EGR1-responsive luciferase construct and a constitutively expressing Renilla construct (40:1). The EGR1-responsive luciferase construct encodes the firefly luciferase reporter gene under the control of a minimal (m)CMV promoter and tandem repeats of the EGR1 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 EGR1, and hence the activity of the EGR1 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 EGR1 and determine the effect of various treatments, such as gene knockdown, over-expression, and chemical compounds on EGR1 activity.

Using a simple dual-luciferase assay, the EGR1 Reporter (luc) can easily and rapidly:

Monitor EGR1 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

293-H cells were transfected with EGR1 reporter, negative control and positive control (for transfection protocol, please refer to the user manual). After 4 hours of transfection, medium was changed to growth medium. After 24 hours of transfection, medium was changed to assay medium (Opti-MEM + 0.5% FBS + 0.1mM NEAA + 1mM Sodium pyruvate + 100 U/ml penicillin + 100 µg/ml streptomycin). After 26 hours of transfection, cells were treated with PMA (10 ng/mL) for 6 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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