LightSwitch™ Synthetic Response Elements

Synonyms

LightSwitch, Light Switch, GoClone, Go Clone, reporter

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Overview

LightSwitch™ Synthetic Response Element reporter constructs simplify the task of determining the effect that test compounds and/or conditions have on over 90 different transcription factors. Each synthRE vector contains an optimized synthetic response element, made up of repeats of a transcription factor binding site motif, cloned upstream of a minimal promoter and the RenSP luciferase gene in the Long-range Enhancer Reporter Vector, pLightSwitch_LR vector. These transfection-ready LightSwitch GoClones^® together with the highly optimized assay reagents, transfection reagents, controls and detailed protocols allows you to perform your transcription factor reporter assay experiments immediately.

Note: All LightSwitch constructs MUST be used with the LightSwitch Luciferase Assay Reagents to obtain optimal results with our novel RenSP luciferase. We also recommend that you include appropriate positive & negative control vectors when you perform your assays.

Description

Each Synthetic Response Element reporter construct includes an optimized synthetic response element that is made up of multiple repeats of a transcription factor binding site motif. In contrast to our LightSwitch Promoter Reporter vectors, which contain endogenous human promoters, Synthetic Response Element vectors contain the synthetic element cloned upstream of a minimal HSV-TK promoter and the RenSP luciferase reporter gene in the pLightSwitch_LR vector. This makes the empty pLightSwitch_LR vector ideal for use as a negative control, as it contains only a basal promoter. For complete information on the pLightSwitch_LR vector, please see the Empty LightSwitch™ Reporter Vectors page.

Data

AR Synthetic Response Element (Androgen Receptor)

HT1080 fibrosarcoma cells were co-transfected with the AR synthRE reporter construct (Cat. No. 32107) and an AR cDNA construct according to our standard protocols. Half of the transfected cells were treated with 10 nM R1881 for 24 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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CREB Synthetic Response Element

HT1080 fibrosarcoma cells were transfected with the CREB synthRE reporter construct (Cat. No. 32117) according to our standard protocols. Half of the transfected cells were treated with 100 nM PMA for 4 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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ER Synthetic Response Element (Estrogen Receptor)

HT1080 fibrosarcoma cells were co-transfected with the ER synthRE reporter construct (Cat. No. 32125) and an ER cDNA construct according to our standard protocols. Half of the transfected cells were treated with 10 nM β-estradiol for 24 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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GR Synthetic Response Element (Glucocorticoid Receptor)

HT1080 fibrosarcoma cells were transfected with the GR synthRE reporter construct (Cat. No. 32140) according to our standard protocols. Half of the transfected cells were treated with 100 nM dexamethasone (DMSO) for 4 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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HIF-1 alpha Synthetic Response Element (Hypoxia)

HT1080 fibrosarcoma cells were transfected with the HIF-1 alpha synthRE reporter construct (Cat. No. 32141) according to our standard protocols. Half of the transfected cells were treated with a low oxygen condition (1% O₂) for 24 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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HSF Synthetic Response Element (Heat Shock)

HT1080 fibrosarcoma cells were transfected with the HSF synthRE reporter construct (Cat. No. 32144) according to our standard protocols. The transfected cells were then incubated at 37°C or 43°C for 8 hours. The cells were then assayed using the LightSwitch Luciferase Assay Kit.

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NFκB Synthetic Response Element (Inflammation)

HT1080 fibrosarcoma cells were transfected with the NFκB synthRE reporter construct (Cat. No. 32160) according to our standard protocols. Half of the transfected cells were treated with 50 ng/ml TNF-α for 8 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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STAT1 Synthetic Response Element (Inflammation)

HT1080 fibrosarcoma cells were transfected with the STAT1 synthRE reporter construct (Cat. No. 32183) according to our standard protocols. Half of the transfected cells were treated with 500 U/ml Interferon-α for 8 hours. The untreated and treated cells were then assayed using the LightSwitch Luciferase Assay Kit.

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