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How can I remove the cells from the 100 uL Nucleocuvette® Vessel after Nucleofection®? Is there any alternative?

We recommend using the plastic single use pipettes provided in the kits. These pipettes prevent any damage to the cells and ensure a good cell recovery.

What is the basic principle of the Nucleofector® Technology?

The Nucleofector® Technology is a novel transfection method especially designed for the needs of primary cells and difficult-to-transfect cell lines. It is a non-viral method based on a unique combination of electrical parameters and cell-type...

Why is the Nucleofector®Technology ideal for primary cells and difficult-to-transfect cell lines?

There are several reasons to choose the Nucleofector® Technology for your gene transfer experiments. One is the direct transfer of DNA to the cell nucleus. This makes gene expression independent from cell division. Therefore, the technique is the...

What optimization is necessary to get the Nucleofector® Technology to work?

None. For each cell type in our product list we offer a cell-type specific solution and thoroughly optimized electrical parameters. These are already pre-programmed in the Nucleofector® Device. Additionally our optimized protocols give you...

What can I do if there is no Nucleofection® optimized protocol available for my cell of interest?

Lonza is constantly developing new optimized kits and protocols for an increasing number of primary cells and cell lines. In order to obtain the latest information, check our website or contact our Scientific Support Team.We are offering Primary Cell...

What are the requirements for direct Nucleofection® of mRNA for protein expression?

The mRNA should be capped and polyadenylated. The conditions of Nucleofection® will be the same as for DNA with the particular cell type, i.e. follow the same protocol and use the same program, except one will likely need to add a much higher...

Is there a special recommendation on Human T cell enrichment before Nucleofection®?

For Nucleofectio® it is preferable to use Human T cell populations enriched by magnetic separation. For example, MACS™ Microbeads (Miltenyi) or by a rosetting method. For magnetic separation we recommend negative selection or detaching beads...

Are there any recommendations to avoid excessive mortality during T cell Nucleofection®?

Try to keep the DNA amount for Nucleofection® quite low (e.g. 1 µg plasmid DNA per 100 µl reaction) since higher DNA amounts might cause increasing toxicity and mortality. It is also important to use highly purified DNA, e.g. by using Endofree™...

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

DNA amount and quality are very critical for Nucleofection® of Dendritic Cells (recommended 0.5-1 µg; maximal 2 µg). LPS (lipopolysaccharides) have a strong negative effect on cell viability. Please make sure that all components you use for dendritic...

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...
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