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673 results sorted by
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GFP expression has been observed for up to one week as measured by light and fluorescence microscopy.
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In general, all reporter genes can be used. For some applications (e.g. reporter gene experiments in various primary cells, experiments with late analysis time point due to starving and /or stimulation) a reporter gene with a longer half life than...
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The overall amount of 5 µg should not be exceeded. Look in the cell specific Optimized Protocol (OP) for the recommended total DNA amount of the cell type of interest.
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Insect cells are cultivated at lower temperatures than E.coli or mammalian cells (S2 cells at 24°C, Sf9 cells at 28°C). Therefore they need a dedicated incubator with appropriate temperature.
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The time course of plasmid expression after Nucleofection™ might be different from the time course seen with other transfection methods. We recommend looking at different time points and start with the first time point as early as 4 to 6 hours post...
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In general the overall expression level differs between different cell types. Don't expect the same level of gene induction when working with primary cells. You might need to change to other DNA amounts or ratios.
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Vectors with a CMV promoter [e.g. pmaxCloning™, Lonza, Cat. No. VDC-1040] typically work fine for transient protein expression in mammalian cells, like HEK293, CHO or NSO. For insect cells, like Sf9 or Sf21, vectors with insect promoters are...
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The optimal incubation time ranges from 2-days to 6 days depending on various factors like the stability of the protein in the medium, potential toxicity of the protein, cell density and stability of the plasmid in the nucleus. To check the...
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For many cell lines in suspension, especially sCHO, conventional transfection methods are very limited in efficiency. For protein production, suspension cells are favored over adherent cells because they are much easier to cultivate. Unlike...
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The generation of a stable clone often requires six months or more due to selection procedures and adaption to serum-free conditions after transfection. With the help of the Nucleofector technology suspension cells can be transfected directly in a...
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