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My suspension-adapted HEK293 (or CHO) cells tend to clump in culture. How can I avoid this?

Efficient protein production experiments require growth of the host cell in single-cell suspension, something that is sometimes difficult to achieve since most established cell lines retain their adherent growth characteristics. Commercially...
Missing: stable linesGuideline

When I transfected my cells, error 8 appeared on the display of the Nucleofector® IIN, IIS or 2b. What is the reason?

...to the incomplete delivery; however, the cells can still be plated for analysis. This generally occurs because the conductivity of the solution was outside of accepted limits – this can happen if the media was not completely aspirated before resuspending, too much...
Missing: stable linesGuideline

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...
Missing: stable linesGuideline

How much sample do you need for accurate results with MycoAlert® Assay? Do you need cells or just supernatant?

The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.
Missing: stable linesGuideline

Can I use the ToxiLight™ Non-Destructive Cytotoxicity BioAssay Kit to assess the effect of transfection procedures on the health of my cells?

...Kinase are present in your transfected cells culture supernatant indicating that they have been damaged by the transfection procedure. This would impact on your transfection efficiency values...
Missing: stable linesGuideline

I tested my cell culture supernatant negative for mycoplasma contamination using the MycoAlert® Mycoplasma Detection Kit or the MycoAlert® PLUS Mycoplasma Detection Kit. But when I tested the same cells using a PCR-based Assay, I obtained a positive result.

It is possible that the PCR is picking up dead mycoplasma whereas the MycoAlert® and MycoAlert® Plus Assay will pick up only viable mycoplasma. It is possible of course that there is contamination in the PCR giving a false positive.
Missing: stable linesGuideline

What cell number do I need per well when using the 96-well Shuttle® Device or the 4D-Nucleocuvette® Stripes?

The cell number is very much dependent on the cell type you use. For high throughput Nucleofection®, we usually start with one fifth of the optimal amount recommended for the standard Nucleofection® in the 100µl cuvette. Currently the absolute...
Missing: stable linesGuideline

In your optimized Nucleofection® Protocols for MDA-MB-231 and MDA-MB-468, you say to culture the cells without CO2. Why?

The media recommended for these cells is Leibovitz's L-15 medium which does not contain sodium bicarbonate to act as a buffer for the carbonic acid that would normally form in the presence of CO2. Furthermore, if the cells are cultured in Leibovitz...
Missing: stable linesGuideline

Do we have any Diseased Human Bronchial/Tracheal Epithelial Cells (DHBE) screened for Air Liquid Interface (ALI) differentiation using the B-ALI™ BulletKit™?

Yes. All our DHBEs are also offered in a version screened for ALI differentiation. The lots which do pass this additional ALI testing are electronically designated by adding an “S” to the end of the original catalog number.
Missing: stable linesGuideline

Do you know if treating cells with 10mM MgCl2 would interfere with the ToxiLight™ Non-Destructive Cytotoxicity BioAssay?

Magnesium (Mg) is a cofactor for both luciferase and Adenylate Kinase (AK) and will speed up their reactions, but this amount of MgCl2 wouldn't make a significant difference.
Missing: stable linesGuideline
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