Data Type

Category

+ Show All

Research Area

+ Show All
684 results sorted by

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,...

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...

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)...

Can I order any of the Nucleofector™ Kit components separately?

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

Who is the distributor for my country?

Please click on the link provided to search for your distributor.

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...

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 are the critical steps for successful Nucleofection™ of monocytic cell lines like THP-1, HL60 and U937?

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...

Do you recommend a particular supplier for siRNA duplexes for use in Nucleofection™ ?

For siRNA applications using the Nucleofector technology we recommend siRNA duplexes from Dharmacon, part of Thermo Fischer Scientific.

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.
PAGE 4

Privacy | Legal | About Lonza

©2019 Lonza. All rights reserved.