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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.

My Nucleofector Solution was frozen. Is this a problem?

As long as the supplement was not added to the Nucleofector™ Solution, then there is no risk of any damage to the solutions. Even long-term storage of several months did not alter the performance of the Nucleofector™ Solution. However, if the...

Is it possible to use different substrates (e.g. siRNA duplexes) with the 96-well Shuttle Device?

Yes. Transfection can be done with any substrate AND under exactly the same conditions. For example, the same conditions (96-well Shuttle™ Solution and program) are used for siRNA transfection or shRNA vectors. This greatly facilitates the...

I have a GFP construct that I use and usually get anywhere from 50-80% GFP positive cells using FACS analysis while I get only 30-40% using a GFP-NFAT fusion protein driven by the same promoter. Do you have any suggestions?

The Amaxa™ Nucleofection™ Conditions are optimised for each cell type, the solutions and programs are cell type dependent and not vector dependant. The expression of the fusion protein depends on the localization of the protein transcription,...

What is the advantage of the 96-well Shuttle Device over common lipofection, especially in a high throughput framework?

Several primary cells of high research relevance can simply not be transfected at relevant efficiencies and viabilities using lipofection. The same holds true for many suspension cell lines which are often used for protein production. These difficult...

What are the contents of a 96-well Shuttle Kit?

As the system will most likely be used for high-throughput approaches we provide plates which are pre-equipped with 96-well Nucleocuvette™ modules (2x8). In addition to that, the 96-well Shuttle solution, the supplement, and the pmaxGFP™ are also...

I currently work with some adherent cell lines, such as 293, which are grown in DMEM media. I have noticed that the mortality seems to be high after Nucleofection. Do you have any recommendations?

For some adherent cell lines we strongly recommend using RPMI instead of DMEM medium. As described in culture conditions of the cell line optimization kit protocol as well as the NIH3T3 (DSMZ) protocol, using RPMI instead of DMEM as "transfer and...

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...

Can I use the Nucleofector Technology for RNAi applications? How do I start?

Yes. The Nucleofector™ Technology can be easily applied for any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the cell type specific Optimized Protocol for DNA vectors (cDNA, shRNA or miRNA expressing plasmids) or...

Can I order any of the Nucleofector Kit components separately?

No. The Nucleofector™ Kits are only available as full kits.
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