SRE Luciferase Reporter Lentivirus

Catalog #

78627

$835 *

Size: 500 µl x 2

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*US Pricing only. For international pricing, please contact your local distributor.

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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 (Figure 1). After transduction, activation of the MAPK/ERK signaling pathway in the target cells can be monitored by measuring the luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the SRE Luciferase Reporter Lentivirus.

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Product Info

Storage and Usage

Citations

Supplied As

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

Materials Required But Not Supplied

Name	Ordering Information
HEK293 cells	ATCC #CRL-1573
Thaw Medium 1	BPS Bioscience #60187
Assay Medium 1B	BPS Bioscience #79617
Recombinant human EGF	BPS Bioscience #90201-1
FBS	Invitrogen #26140-079
Polybrene	Millipore, #TR-1003-G
96-well tissue culture, clear-bottom, white plate	Corning, #3610
One-Step Luciferase assay system	BPS Bioscience #60690
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Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.

Background

The MAPK/ERK signaling pathway is a major participant in the regulation of cell growth and differentiation. It can be activated by various extracellular stimuli including mitogens, growth factors, and cytokines. Upon stimulation, MEK1/2 phosphorylate and activate ERK1/2. The activated ERK1/2 translocate to the nucleus where they phosphorylate and activate transcription factors. The TCFs (Ternary Complex Factors), including transcription factor Elk1, are among the best-characterized substrates of ERK. When phosphorylated by ERK, Elk1 forms a complex with Serum Response Factor (SRF) and binds to the Serum Response Element (SRE), resulting in the expression of numerous mitogen-inducible genes.

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.

Growth Media

For best results, it is highly recommended to use these validated and optimized media from BPS Bioscience. Other preparations or formulations of media may result in suboptimal performance.

Thaw Medium 1 (BPS Bioscience, #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Assay Medium 1B (BPS Bioscience, #79617): MEM medium supplemented with 0.5% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Applications

Screen for activators or inhibitors of MAPK/ERK signaling pathway in transduced target cells
Generate SRE Luciferase Reporter stable cell lines (puromycin resistant)

Shipping Temperature

-80°C

Notes

Biosafety
The lentiviruses are produced with a third generation SIN (self-inactivation) lentivector which
ensures self-inactivation of the lentiviral construct after transduction and after 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 and are not present in the lentivirus particle. 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.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze-thaw cycles.

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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SRE Luciferase Reporter Lentivirus

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