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How can I improve Human Hepatocyte viability post Nucleofection™ on the 96-well Shuttle™ device?

In addition to proper cell handling and using culture media optimized for long term culturing of hepatocytes (Lonza Basal medium supplemented with BulletKit), we recommend separating the healthy hepatocytes from the dead cells and debris in the...

What protocol should I use for Nucleofection® of patient derived blood samples, e.g. leukemia or lymphoma cells?

Unfortunately, we do not have a ready-to-use protocol for Chronic lymphocytic leukemia (CLL) or other blood cell derived cancer cells. The cells often have a chromosomal aberration and show the properties of a cell line. Therefore we suggest to...

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...

Does the GFP sequence in the pmaxGFP™ control plasmid has a stop codon?

Yes. The coding sequence ends with a stop codon (TGA/UGA).

Can mycoplasma grow in the absence of cells?

There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present.

How long is the MycoAlert® Mycoplasma Detection Assay signal stable?

Please follow the protocol exactly as written – add reagent, wait 5 min and read, add substrate, wait 10 minutes and read again.

What kind of effects could a mycoplasma contamination have on my cells?

Mycoplasma contamination can: > Inhibit cell metabolism > Cause cell death > Induce morphological alterations > Alter DNA, RNA, and protein synthesis > Induce chromosomal aberrations > Cause increased sensitivity to inducers of...

Can I also use the Lucetta® 2 Luminometer for other assays besides the MycoAlert® Mycoplasma Detection Assay?

Yes. In addition to the tailor-made MycoAlert® Mode it also has a "Single Read" mode which is suited for unprocessed luminescence readings, e.g. ATP-based cell proliferation or cytotoxicity assays, or Luciferase reporter gene assays.

Is it possible to quantify the mycoplasma with the MycoAlert® Mycoplasma Detection Assay?

No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.

When can I start induction studies on Human Hepatocytes transfected with the Nucleofector®?

We recommend allowing the transfected hepatocytes to recover in Plating medium for 72 hours post-Nucleofection®.For induction studies (e.g. CYP3A4), the Plating medium is replaced with maintenance medium (William's E without FCS and EGF) and...

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