eGFP Lentivirus (Inducible TET On)

Catalog #

78629

$835 *

Size: 500 µl x 2

Qty

Bulk/Custom

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

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Description

Enhanced green fluorescent protein (eGFP) is a modified (F64L and S65T mutations) version of the native GFP protein isolated from jellyfish (Aequorea victoria), displaying increased fluorescence and more efficient folding. Enhanced GFP Lentiviruses (Inducible TET On) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to infect almost all types of mammalian cells, including primary and non-dividing cells. These viruses constitutively express eGFP under a tight TRE tetracycline-inducible promoter (Figure 1). After induction with Doxycycline, eGFP expression can easily be visualized and optimized via fluorescence microscopy or flow cytometry. eGFP has an excitation wavelength of 488 nm, an emission wavelength of 509 nm, and extinction coefficient of 55,000 M^-1cm^-1.

Figure 1. Schematic of the lenti-vector used for the eGFP lentivirus (Inducible TET on). Note that NeoR confers resistance to Geneticin (G418).

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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 16	BPS Bioscience #78647
Doxycycline	Sigma #D9891
Polybrene	Millipore, #TR-1003-G
96-well tissue culture-treated assay plates
Flow cytometer or fluorescence microscope

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture 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.

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. To ensure the maximal range of induction, we recommend using serum which does not contain trace levels of tetracycline (or its derivatives).

Thaw Medium 16 (BPS Bioscience, #78647): MEM medium supplemented with 10% TET-free FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Applications

Ideal as a positive control for transduction with a tetracycline-inducible expression system; useful for transduction optimization.
Generation of a TET inducible stable cell line expressing eGFP with G418 selection.

Shipping Temperature

-80°C

Notes

To generate the TET Inducible Enhanced GFP stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of G418 (as pre-determined from a killing curve) for antibiotic selection of transduced cells.

Biosafety
The lentiviruses are produced with a 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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eGFP Lentivirus (Inducible TET On)

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