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Application: Plasma membrane labelling.Generation of cell pools or stable cell lines expressing membrane 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 mutant variants, such as the eGFP (enhanced GFP). eGFP has a higher intensity emission versus the GFP molecule. Membrane-targeting sequence has been used for artificial localization of green fluorescent protein (GFP) to the membrane.
Description: Membrane 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 eGFP (enhanced green fluorescent protein), with a plasma membrane-targeting sequence at the N-terminus of eGFP, under the control of an EF1a promoter. The lentiviruses also transduce a puromycin selection marker. eGFP expression and transduction efficiency can easily be verified and optimized via fluorescence microscopy or flow cytometry.
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 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 (50 µl x 2) of lentivirus at a titer ≥1 x 108 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
References: s