Psicheck-2 Dual Glo Luciferase Reporter Assay Protocol

Description: The ADAR1 Luciferase Reporter HEK293 cell line is designed to monitor RNA editing by Adenosine deaminase acting on RNA (ADAR1). This cell line stably expresses ADAR1 under the control of a CMV promoter and a separate ADAR editing reporter construct expressed under the control of another CMV promoter. The reporter contains the gene encoding firefly luciferase, which is constitutively expressed in the cells, upstream of the gene encoding the GluA2 ADAR substrate followed by the Renilla luciferase gene. The sequence corresponding to GluA2 has been modified to contain an amber stop codon (UAG). When edited by ADAR, this stop codon (UAG) will be changed to UIG (A to I edit), which is read as tryptophan (UGG) by the translation machinery. This edit allows translation to occur all the way to the end of the reporter mRNA and results in the expression of Renilla luciferase. Conversely, in the absence of ADAR1 activity, translation terminates at the stop codon and Renilla is not expressed. Reporter activity is read out as the Renilla Luciferase/Firefly luciferase ratio whereby inhibition of ADAR activity, and thus the UAG (stop) to UGG (tryptophan) conversion rate, will result in a dose-dependent decrease in the Renilla luciferase/Firefly luciferase ratio.

Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.

Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.

Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.

Description: The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly. This reagent lyses the cells and contains a substrate for firefly luciferase to produce firefly luciferase luminescence. Next, the Renilla Luciferase Reagent is added to the same well. It quenches the firefly luciferase luminescence and provides the substrate for renilla luciferase to produce renilla luciferase luminescence. The light production of both reactions can be conveniently measured on a luminometer. This assay system has several features:• Sensitive: highly sensitive detection of firefly luciferase activity and Renilla luciferase activity.• Stable: the luciferase signal output is stable for more than one hour, providing flexibility with regard to incubation time• High-throughput: simple homogeneous protocol minimizes handling steps to support high-throughput screening applications• Compatibility: works well with a variety of common media containing 0-10% serum and phenol red.

Description: The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly. This reagent lyses the cells and contains a substrate for firefly luciferase to produce firefly luciferase luminescence. Next, the Renilla Luciferase Reagent is added to the same well. It quenches the firefly luciferase luminescence and provides the substrate for renilla luciferase to produce renilla luciferase luminescence. The light production of both reactions can be conveniently measured on a luminometer. This assay system has several features:• Sensitive: highly sensitive detection of firefly luciferase activity and Renilla luciferase activity.• Stable: the luciferase signal output is stable for more than one hour, providing flexibility with regard to incubation time• High-throughput: simple homogeneous protocol minimizes handling steps to support high-throughput screening applications• Compatibility: works well with a variety of common media containing 0-10% serum and phenol red.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase.

Description: The IL-2 Promoter Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the human IL-2 promoter. After transduction, activation of the IL-2 signaling pathway in the target cells can be monitored by measuring the luciferase activity._x000D_

Description: The Nrf2 antioxidant response pathway plays an important role in the cellular antioxidant defense. Nrf2, a basic leucine zipper transcription factor, induces the expression of antioxidant and phase II enzymes by binding to the ARE (antioxidant response element) region of the gene promoter. Under basal conditions, Nrf2 is retained in the cytosol by binding to the cytoskeletal protein Keap1. Upon exposure to oxidative stress or other ARE activators, Nrf2 is released from Keap1 and translocates to the nucleus, where it can bind to the ARE, leading to the expression of antioxidant and phase II enzymes that protect the cell from oxidative damage.
The ARE Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by ARE located upstream of the minimal TATA promoter. After transduction, activation of the Nrf2 antioxidant response pathway in the target cells can be monitored by measuring the luciferase activity.

Description: The SRE (Serum Response Element) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the Serum Response Element located upstream of the minimal TATA promoter . After transduction, activation of the MAPK/ERK signaling pathway in the target cells can be monitored by measuring the luciferase activity.

Description: The Myc Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the Myc response element located upstream of the minimal TATA promoter and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the Myc signaling pathway in the target cells can be monitored by measuring the luciferase activity.

Description: The UAS (Upstream Activation Sequence) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by a multimerized GAL4 upstream activation sequence (UAS) located upstream of the minimal TATA promoter and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the UAS-controlled signaling pathway in the target cells can be monitored by measuring the luciferase activity.

Description: The p53 Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by p53 response elements located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, p53-regulated gene expression in the target cells can be monitored by measuring the luciferase activity.

Description: The Hypoxia Response Element (HRE) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by four copies of a hypoxia response elements (HRE) located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the induction of hypoxia in the target cells can be monitored by measuring the luciferase activity.

Description: Human IL-2 reporter construct is stably integrated into the genome of Jurkat T-cells. The firefly luciferase gene is controlled by a human IL-2 promoter.