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View additional product information for CytoTune™ EmGFP Sendai Fluorescence Reporter - FAQs (A16519)
7 product FAQs found
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.
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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.
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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).
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
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.
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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.
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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.
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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.