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DNA amount and quality are very critical for Nucleofection™ of Dendritic Cells (recommended 0.5-1µg; maximal 2 µg). LPS (lipopolysaccharides) have a strong negative effect on cell viability. Please make sure that all components you use for dendritic...
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The PBS should be calcium and magnesium free to prevent clumping of cells. We routinely use PBS without calcium and magnesium for example Lonza Cat. No. 17-516F
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After Nucleofection™, wait at least four hours before stimulation. Stimulating immediately after Nucleofection™ may lead to increased cell mortality and poor activation of cells. For more details on the stimulation of Human T cells post...
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The pmaxGFP control DNA is not suitable for making stable clones because it lacks a selectable marker for mammalian cells. The kanamycin resistance expressed by this plasmid is only suitable for bacterial selection.
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The correct rotor speed can be calculated by measuring the maximum radius of your rotor and entering the information into the table found on the website which can be reached by clicking on the Related Link.
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The expiration dates are located on the outside label of the kit box and on the label of the solution box.
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Yes. We recommend using mice between 6-12 weeks. Using mouse T cells isolated from younger or older animals for Nucleofection™ may result in much lower transfection efficiencies and/or viabilities.
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?
No. The post Nucleofection™ Recovery media contains no immunogenic factors and should not influence cell stimulation or differentiation. For Mouse T cells, some experiments have shown that after Nucleofection™ cells can be stimulated efficiently in...
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?
We have examined the effects of Nucleofection™ (without DNA) on these cells and have not observed significant activation. This indicates that neither the Nucleofector™ Solution, the Nucleofector™ Program nor the recovery medium are sufficient for...
M
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As long as the supplement was not added to the Nucleofector™ Solution, then there is no risk of any damage to the solutions. Even long-term storage of several months did not alter the performance of the Nucleofector™ Solution. However, if the...
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