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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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No. The 4D-Nucleofector and 96-well Shuttle™ System uses specifically optimized electrical parameters and solutions. Five different 4D-Nucleofector or 96-well Shuttle™ Solutions are available for primary cells. For cell lines, three different 96-well...
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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 timepoint...
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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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The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.
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Antibiotics should be added 24-48 h after Nucleofection.
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According to QBM Cell Sciences, you can expect 12 million cortical neurons from an average E18-E19 rat pup. You can expect 4 million cortical neurons from an average mouse pup.
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Every 2-3 days. This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.
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This depends on the respective cell type (transfection efficiency, viability and integration frequency). If it is important to you to know the frequency of stable integrants before carrying out your actual experiments, the frequency should be...
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