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Do you have any siRNA Nucleofection results using concentrations lower than 50nM?

Yes. Please click on the attached Technical Reference Guide: "Designing an RNAi Experiment Using Nucleofection™". On page 3 you can find a table with some examples.

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 non-dividing...

The animal facility which isolates my hepatocytes is at a different location, should I be concerned about transport time?

Isolated hepatocytes can be transported in Krebs-Henseleit Buffer or supplemented William's E Medium. Long transport time for isolated hepatocytes, (in excess of one hour) can cause a decrease in efficiency of Nucleofection™, but does not seem to...

How much DNA can I use when transfecting hepatocytes with the standard Nucleofector?

We have used up to 6 µg of DNA per 100 µl reaction with no deleterious effect.

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.

For hepatocytes, is plating density a concern post Nucleofection?

Yes, the optimized protocols for the standard Nucleofector™ recommend the use of 6 well plates. Plating into a 12-well plate will pose a problem as less cells will attach at high densities.

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....

Four hours after Nucleofection, I can see the hepatocytes have attached to the well, but the morphology does not look correct, should I be concerned?

There is no reason for alarm. The hepatocytes may not exhibit normal morphology a few hours after Nucleofection™ but by 24 hours post Nucleofection™, morphology should be normal. Remember to perform a fluid change about 4 hours post Nucleofection™....

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...

Can calcium influx issues affect viability of human Mesenchymal Stem Cells (hMSC)?

Yes. When cells are transfected using Nucleofection™, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection™ in media that...
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