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Application: Sub-cellular (Nuclear) labelling of cellsGeneration of cell pools or stable cell lines expressing firefly luciferase and nuclear eGFP following puromycin selection
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 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. The presence of firefly luciferase allows for easy assay read-outs using luminescence.
Description: The Firefly Luciferase-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 firefly luciferase and nuclear eGFP (Luc2-P2A-nuclear eGFP) driven by an EF1A promoter. These lentiviruses also transduce a puromycin selection marker (Figure 1). The nuclear eGFP has NLS sequences at the C-terminus. eGFP has an excitation wavelength of 488 nm, an emission wavelength of 509 nm, and extinction coefficient of 55,000 M-1cm-1.
Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture 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.
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.
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.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
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