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Invitrogen™ CytoTune™ EmGFP Sendai Fluorescence Reporter

Catalog No. A16519
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Catalog No. A16519 Supplier Invitrogen™ Supplier No. A16519
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Description

The CytoTune™ EmGFP Sendai Fluorescence Reporter helps you determine whether the Sendai particles that are used in all CytoTune™-iPS Sendai reprogramming kits can transduce into a given cell type before you advance to one of these kits.

This vector expresses the next-generation Aequorea victoria EGFP gene, encoding the emerald green fluorescent protein (EmGFP). The cell will express the EmGFP, which is bright, photostable, and ideal for testing transduction.

The CytoTune™ EmGFP Sendai Fluorescence Reporter enables a better understanding of how both the original CytoTune™-iPS Sendai Reprogramming kits and the newer CytoTune™-iPS 2.0 Sendai Reprogramming Kit transduces various cell types. The Sendai particle enters a cell via its HN envelope protein that recognizes sialic acid and is widely expressed in mammalian cells, allowing Sendai virus to target a wide range of cell types. The Sendai virus vectors do not enter the nucleus, thus avoiding any DNA phase, as the viral genome remains as RNA in the cytoplasm.

Specifications

Product Type Sendai Reporter
Gene GFP (EmGFP)
Assay GFP (EmGFP)
Content And Storage 1 Tube, store (at -68°C to -85°C.)
Delivery Type Sendai virus
Promoter Proprietary
Product Line CytoTune
Quantity 1 Pack
Form Liquid

Frequently Asked Questions (FAQs)

Can I mix the Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter with the Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit?

If you want to use the EmGFP Reporter with reprogramming, it must be added at the time of reprogramming. Cells infected with Sendai virus will most likely be refractive to further infection. Therefore, do not try to add Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit to cells already transduced with Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter or vice versa.
After addition of Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter, when can I see expression of EmGFP?

The expression of EmGFP in successfully transduced cells is detectable at 24 hours post-transduction by fluorescence microscopy and reaches maximal levels at 48-72 hours post-transduction.
How do I use the Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter?

Add the reporter one time to your cells and monitor for expression. Different cell types may vary in their ability to take up Sendai virus; therefore, we suggest initially transducing your cells with at least 2-3 different MOIs (e.g., 1, 3, and 9). Please refer to the user manual for the full protocol (http://tools.thermofisher.com/content/sfs/manuals/cytotune_emgfp_sendai_reporter_man.pdf).
What is EmGFP?

Emerald Green Fluorescent Protein (EmGFP) is a form of GFP with brighter green fluorescence, used to report the expression of a gene of interest. EmGFP can be visualized on standard FITC channel.

What is the Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter?

The Invitrogen CytoTune-EmGFP Sendai Fluorescence Reporter is a fluorescent control vector carrying the EmGFP gene. The fluorescent control vector allows the determination of whether a cell line of interest is amenable or refractive to infection by Sendai reprogramming vectors.

When should I use the full length Human FGF-basic (FGF-2/bFGF) Recombinant Protein or the truncated variant, Human FGF-basic (FGF-2/bFGF) (aa 10-155) Recombinant Protein?

The full length Human FGF-basic (FGF-2/bFGF) (aa 1-155) Recombinant Protein is recommended for stem cells whereas the truncated variant, Human FGF-basic (FGF-2/bFGF) (aa 10-155) Recombinant Protein which is missing the first 9 amino acids, is recommended for use with neural and cardiac cells.
What are induced pluripotent stem cells?

Induced pluripotent stem cells (iPS or iPSCs) are pluripotent stem cells directly generated by introducing combination of genes coding for “reprogramming factors” into adult cells. These reprogramming factors include Oct4, Sox2, c-Myc, KLF4, NANOG, and LIN28. Yu, et al, generated iPS from a human mesenchymal cell line using lentiviral vectors carrying Oct4, Sox2, NANOG, and LIN28 genes (Science 318:1917 (2007)). Using a similar approach, Takahashi et al, generated iPS from human primary fibroblast cells by introducing genes coding for Oct3, Sox2, KLF4, and c-Myc into these cells (Cell 131:861 (2007)). iPS generated by reprogramming are similar to human ES cells in morphology, the capacity for unlimited proliferation, surface-antigen expression, gene expression, the ability to differentiate into cell types representing the three germ layers in vitro, and the ability to form teratomas after injection into SCID mice.

For Research Use Only. Not for use in diagnostic procedures.