Nuclear eGFP Lentivirus (Puromycin)

Catalog #

78976

$875 *

Size: 500 µl x 2

Qty

Bulk/Custom

Add to Favorites

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

●

Purchase

●

Description

Nuclear eGFP Lentivirus 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. These particles contain a nuclear eGFP (enhanced green fluorescent protein), with nuclear localization sequences (NLS) at both N and C-terminus of eGFP), under the control of an EF1a promoter. The lentiviruses also transduce a puromycin selection marker (Figure 1). eGFP expression and transduction efficiency can easily be verified and optimized via fluorescence microscopy or flow cytometry. eGFP has an excitation wavelength of 488 nm, an emission wavelength of 509 nm, with an extinction coefficient of 55,000 M^-1cm^-1.

Figure 1. Schematic of the lenti-vector used to generate the Nuclear eGFP Lentivirus

●

Product Data Gallery

eGFP expression in HEK293 cells transduced with Nuclear eGFP Lentivirus.

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
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
96-well tissue culture-treated assay plates
Flow cytometer or fluorescence microscope

Formulation

The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.

Background

GFP (green fluorescent protein) presents green fluorescence, and it was first identified in Aequorea Victoria. It has become widely used in cell biology to monitor gene expression, protein localization, and protein-protein interactions. Its popularity prompted the development of mutant variants, such as the eGFP (enhanced GFP). eGFP has a higher intensity emission versus the GFP molecule. Nuclear localization signal (NLS) sequences have been used for artificial localization of green fluorescent protein (GFP) in the nucleus.

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.

Applications

Nuclear labelling.
Generation of cell pools or stable cell lines expressing nuclear eGFP following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a Nuclear eGFP expressing stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection.

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

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

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

Nuclear eGFP Lentivirus (Puromycin)

PRODUCTS

SERVICES

RESOURCES

COMPANY