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Why do you have different Optimized Protocols for the Nucleofection® of unstimulated and stimulated Human T cells?

We found that the stimulation of T cells not only changes the cells' function but also significantly alters the requirements for successful Nucleofection®. That's why Lonza developed two Optimized Protocols for transfection giving different...

What antibodies can be used against the GFP produced by pmaxGFP™ and pmaxFP™-Green vectors? What antibodies can be used to detect denatured GFP? What antibodies can be used to detect the RFP expressed by pmaxFP-Red?

Most of the commercial antibodies for GFP do not detect GFP expressed from pmaxGFP or pmaxFP-Green vectors. The origin of the protein expressed by the pmaxGFP control plasmid and pmaxFP-Green is from the green fluorescent protein CopGFP, cloned from...

Can I use a Nucleofector® I or Nucleofector® 2b Device together with the 96-well Shuttle® Device?

No, the 96-well Shuttle® only works in conjunction with the Nucleofector® IIs Device or our 4D-Nucleofector® Device.

Is it possible to use frozen primary CD34+ hematopoietic stem cells for Nucleofection®?

Yes. For cryopreserved PBMCs or enriched CD34 populations, we recommend culturing the thawed cells 1-2 hours in culture medium before Nucleofection®. Any further enrichment procedure after thawing is not recommended. Viability of frozen nucleofected...

What is the advantage of the Nucleofector® Technology over common electroporation (systems)?

High cell viability and high transfection efficiency with the Nucleofection® Technology are the most prominent features. Nucleofection®, i.e., the transfer of DNA into the nucleus and not only the cytosol, is essential when working with...

What are the critical steps for successful Nucleofection® of monocytic cell lines like THP-1, HL60 and U-937?

To achieve optimal results, we strongly recommend following the Optimized Protocols in regard to cell culture conditions (medium, splitting cycle, density before Nucleofection), DNA amount and purity.Please also be sure not to exceed 90xg when...

Is it possible to pre-plate substrates when using the 96-well Shuttle® for Nucleofection®?

Yes, pre-plating of substrates like siRNA and subsequent storage at -20°C is possible.

When nucleofecting cancer cell lines, do I need to be concerned about passage number?

For the most efficient gene transfer, we recommend using cells that are in logarithmic growth phase and at a passage number of 10-15 (from the time of thaw). This is because some cell lines differentiate and change their features after many...

What kind of protocol you recommend for stimulation of human T cells?

For getting the results we present in our datasheet, we recommend the follwing strategy: Cells should be stimulated and cultivated for 3 days before Nucleofection, preferentially on plates coated with 1 µg/ml anti-CD3 and 2 µg/ml anti-CD28 antibody....

Are there any effects on the differentiation of neural stem cells following Nucleofection®?

The ability of transfected rat or mouse neural stem cells to be subsequently differentiated into a variety of cell types is well documented in:Olig2 Overexpression Induces the In Vitro Differentiation of Neural Stem Cells into Mature...

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