ARE Luciferase Reporter Lentivirus

Catalog #

79869

$875 *

Size: 500 µl x 2

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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 (Figure 1). After transduction, activation of the Nrf2 antioxidant response pathway in the target cells can be monitored by measuring the luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the ARE luciferase reporter lentivirus

This product has been cited 16 times.

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Synonyms

ARE cell line, Nrf2 reporter, Nrf, Hepg2, ARE

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Product Info

Storage and Usage

Citations16

Supplied As

Two vials (500 μl x 2) of ARE luciferase reporter lentivirus at a titer 1 x 10⁷TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

• DL-Sulforaphane (Sigma, #S4441). Prepare 10 mM stock solution in DMSO.
• HepG2 growth medium or use Thaw Medium 9 (BPS Bioscience #79665) (MEM medium (Hyclone #SH30024.01)
supplemented with 10% FBS (Thermo Fisher, Cat. #26140079), 1% non-essential amino acids (Hyclone #SH30238.01), 1 mM Na pyruvate (Hyclone #SH30239.01).)
• Polybrene (Millipore, #TR-1003-G)
• 96-well tissue culture treated white clear-bottom assay plate (Corning, #3610)
• ONE-Step™ luciferase assay system (BPS Bioscience, #60690)
• Luminometer

Formulation

The lentiviruses were produced from HEK293T cells in the medium containing 90% DMEM + 10% FBS.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.

Shipping Temperature

-80°C

Warnings

Biosafety

The lentiviruses are produced with the third generation SIN (self-inactivation) lentivector which
ensures self-inactivation of the lentiviral construct after transduction and integration into the
genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the
transduced cells, as they are expressed from packaging plasmids lacking the packing signal.

Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a
Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and
institutional regulations and using all appropriate safety precautions.

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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ARE Luciferase Reporter Lentivirus

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