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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...

Can calcium influx issues affect viability or differentiation in Stem Cells after Nucleofection?

When cells are transfected by 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 medias that contain high...

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.

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...

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 amount...

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.

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.

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™.

Do you have a protocol for the Nucleofection of Plasmodium berghei?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector™ Solution for your...

Do you have a protocol for the Nucleofection of Plasmodium yoelii?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector Solution for your...
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