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Application: Nuclear labelling.Generation of cell pools or stable cell lines expressing Nuclear mCherry following puromycin selection.
Background: mCherry is a monomeric red fluorescent protein derived from DsRed found in the sea anemones Discosoma. It belongs to the mFruit family of monomeric red fluorescent proteins, which are improved versions of mRFP1 (monomeric red fluorescent protein 1) in terms of brightness and photostability. The use of fluorescent proteins allows for direct visualization of transfected or transduced cells under a fluorescent microscope or analysis by flow cytometry. The use of lentiviruses to introduce mCherry is a convenient strategy that allows expression of the markers in almost all mammalian cells and to easily determine transduction efficiency. Nuclear localization signal (NLS) sequences have been used for artificial localization of red fluorescent protein (mCherry) to the nucleus.
Description: Nuclear mCherry 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 mCherry, with nuclear localization sequences (NLS) at both the N- and C-terminus of mCherry, under the control of an EF1a promoter. The lentiviruses also transduce a puromycin selection marker. mCherry 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