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What is the lowest amount of Human Hepatocytes that can be used with the 96-well Shuttle® System or the 4D-Nucleofector® X-Unit?

Human hepatocytes do not like to be cultured and transfected at low densities. Therefore, using less than 100,000 cells per Nucleofection® is not recommended.

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 kind of effects could a mycoplasma contamination have on my cells?

Mycoplasma contamination can: > Inhibit cell metabolism > Cause cell death > Induce morphological alterations > Alter DNA, RNA, and protein synthesis > Induce chromosomal aberrations > Cause increased sensitivity to inducers of...

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.

How can I detect mycoplasma contamination of my cells?

We recommend using the 20-minute MycoAlert® Mycoplasma Detection Kit from Lonza. The simple four-step MycoAlert® Assay is a selective biochemical test that exploits the activity of mycoplasmal enzymes. The presence of these enzymes provides a rapid...

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....

How can cells be cured of mycoplasma contamination?

Where contamination has occurred, and the sample absolutely cannot be discarded, the MycoZap™ Mycoplasma Elimination Reagent has been optimized to eliminate mycoplasma with minimal toxic effects on the cells. The MycoZap™ Reagent eliminates...

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...

Do you have any instructions available for using a hemocytometer for cell counting?

Yes. Our technical guideline "Cell Counting and Determination of Viability via Hemocytometer" can be found as associated pdf file in the link below.

Do you have data about the in vivo implantation of hepatocytes transfected by Nucleofection? For how long could protein expression be detected?

There is a publication reporting on the successful reinjection of modified hepatocytes into mice.Chen et al. were able to monitor the expression over several days. This group worked with a preliminary test solution for mouse hepatocyte as the...
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