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How can I increase cell viability of Dendritic cells after Nucleofection?

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...

Does it matter if the PBS used for monocyte enrichment contains calcium and magnesium, if the cells should be used later in Nucleofection ?

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

I want to stimulate Human T cells post Nucleofection. Are there any precautions to keep in mind?

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...

Is it possible to get neomycin resistant clones after Nucleofection with pmaxGFP?

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.

How can I determine the correct centrifugation speed for my particular centrifuge and rotor?

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.

Where can I find the expiration dates for the Nucleofector Kit and the Nucleofector Solution?

The expiration dates are located on the outside label of the kit box and on the label of the solution box.

Is the age of the mice important for my mouse T cell Nucleofection?

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.

Does the recovery medium contained in the Mouse T Cell Nucleofector Kits contain any immunogenic substances?

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...

I see activation of my monocytes (or macrophages or DCs) following Nucleofection. Why is this? What can I do to address this problem?

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...

My Nucleofector Solution was frozen. Is this a problem?

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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