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pLV-SFFV-human NIS-P2A-eGFP-T2A-Luc2

  • 20 ug (DNA1041) $ 1,150
  • Name of plasmid: pLV-SFFV-human NIS-P2A-eGFP-T2A-Luc2
  • Description: pHR' SIN-SFFV-hNIS-P2A-eGFP-T2A-Luc2 plasmid DNA
  • Amount of DNA: 20 ug
  • Promoter: SFFV spleen focus forming virus promoter for high constitutive gene expression in numerous cell types. Unlike the CMV promoter, SFFV works very well in human, rodent, and other animal cells,
  • Transgenes: Human sodium iodide symporter (hNIS), enhanced green fluorescent protein (eGFP), and firefly luciferase (Luc2)
  • Linkers: Self-cleavage peptides P2A and T2A
  • Proposed Use: For generation of lentiviral vector stocks encoding human NIS, enhanced GFP, and firefly luciferase in a triple transfection protocol.

Plasmid Map:

plasmid map hNIS eGFP Luc2

Transgene Validation:

Uptake Assay: NIS mediated 125I uptake of LV-hNIS-eGFP-Luc2 transduced cells was assayed in the presence or absence of KClO4, a competitive inhibitor of iodine uptake.

Uptake, hNIS, NIS

Flow Cytometry: LV-hNIS-eGFP-Luc2 transduced (green) or control (grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry.

flow cytometry eGFP

Luciferase Assay: 104, 105, or 106 LV-hNIS-eGFP-Luc2 transduced cells were placed in wells of a 96-well plate and 0.3 mg of d-luciferin was added to the indicated wells. The plate was immediately imaged using a Xenogen IVIS Spectrum.

luciferase assay

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Use Fewer Animals

Our Reduction Campaign

At Imanis, we are committed to promoting the practice of the 3Rs in animal research. Learn how we are decreasing the use of animals and research as well as saving up to 15% on your orders. Continue reading...