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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) or...

Can I order any of the Nucleofector Kit components separately?

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

What method can I use to fix my HL-60 cells as shown in your optimized protocol?

The cells which are shown in the HL-60 optimized protocol are living cells and not fixed cells. However a good method for fixation of GFP is to use 3.5%-4% Paraformaldehyde in PBS. Please keep in mind that the fixation method is dependant on what you...

Who is the distributor for my country?

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

What is the principle behind the ViaLight Plus Cell Proliferation and Cytotoxicity Assay?  

ViaLight™ Plus BioAssay Kits use bioluminescence catalysed by the firefly luciferase enzyme to measure intracellular ATP.  Since ATP is central to the metabolism of all cells, the intracellular concentration is very precisely...

Do ATP levels differ between different cell types?

Healthy cells in culture contain maximal amounts of ATP and a wide range of metabolic stimuli do not increase these ATP levels.  Any increase in the ATP concentration of a culture can therefore be associated with an increase in the number of...

Does the MycoALert® Assay recognize the whole spectrum of mycoplasma?

The mycoplasma specific metabolism detected by the MycoAlert® Assay is present in all members of the mollicute family (Mycoplasma, Acholeplasma, Entomoplasma and Spiroplasma) except Ureaplasma (which are no usual suspects in cell culture). 95% of all...

Can the ViaLight kits be used for both proliferation and cytotoxicity assay

Yes.  Since the ATP levels are precisely maintained in healthy cells, the level of ATP accurately reflects the number of viable cells in a culture.  In response to a proliferative signal (e.g. a growth factor) the number of cells in a...
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