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Applications: Control in experiments that involve transduction.Optimize transduction assays and track transduction efficiency over time.Generate Renilla Luciferase-eGFP reporter cell pools or stable cell lines following puromycin or hygromycin selection.
Background: Renilla Luciferase, also known as RLuc, or Renilla-luciferin 2-monooxygenase, is a bioluminescent protein identified in Renilla reniformis. The chemical reaction that leads to emission of luminescent involves the degradation of colelenterazine to coelenteramide and a photon of blue light. Renilla Luciferase is broadly applied as a reporter in in vitro cell culture and in vivo imaging. 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 interactions. Its popularity prompted the development of mutant variants, such as the eGFP (enhanced GFP). eGFP has a higher intensity emission compared to the GFP molecule. The presence of fluorescent proteins and Renilla Luciferase allows for cell identification and quantification by flow cytometry or fluorescence microscopy, and localization studies in vivo, providing easy assay readouts.
Description: Renilla Luciferase-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. The particles contain Renilla Luciferase and eGFP (green fluorescent protein) (RLuc-P2A-eGFP) driven by an EF1A promoter. The lentiviruses also transduce a puromycin or hygromycin selection marker (Figure 1). The Renilla luciferase and eGFP proteins are under the EF1A promoter and are co-expressed in transduced cells, allowing greater flexibility of detection of the transduced cells.
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.
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 Renilla Luciferase- eGFP Reporter Lentivirus at ≥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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