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I transfected neurons using the 4D-Nucleofector® Y-Unit. After Nucleofection®, there were precipitates in my culture. What can I do?

Most likely there was too much Nucleofector® Solution left before medium addition. Please try to aspirate solution after Nucleofection® as complete as possible without disturbing cell adherence, but add medium quickly to avoid cells getting...

Do I need to use a gelatin, fibronectin, collagen, or Matrigel™ coating when culturing my Clonetics™ Cells?

Most Clonetics™ Cells do not require an extracellular matrix when grown on tissue culture treated plastic – flasks, dishes, well plates (with the exception of hepatocytes, and the rat/mouse neuronal and dorsal root ganglion neurons, rat cardiac...

Your instructions for culturing of Clonetics™ Cells do not mention centrifuging the cells upon thawing. Won't the DMSO kill the cells?

We have seen on multiple occasions that the effects of centrifugation can be significantly more damaging than the effects of residual DMSO on primary cells. As such, unless specifically noted in the recommended culture protocol, instead of...

I thawed and plated my ampoule of Clonetics™ Cells yesterday and most of the cells are floating. What happened?

Clonetics™ Cells will not be confluent on day one. After 24 hours, you will see few attached cells per field under the microscope. Aspirate off the media and the floating cells and replace with fresh, pre-warmed media. Those attached cells will begin...

I've had my Clonetics™ Cells in culture for a few months, they're no longer growing and don't look like healthy cells, what's wrong?

All Clonetics™ Cells are primary cells with a finite life span. Most Clonetics™ Cells are guaranteed for 10 – 15 additional population doublings, which equates to about 2 – 3 passages. As cell death begins to occur, morphology and cell performance...

Do you have any recommendation how to coat glass cover slips for neuron culture, e.g. for use with the Y unit or for post Nucleofection® culture in general?

Preparation of coverslips: At Lonza we used coverslips from Menzel (part of Thermo Scientific). We think that the coverslips have such a good quality that they don’t have to be cleaned prior to use.If you want to use other coverslips or prefer...

What is a Clonetics™ or Poietics™ Basal Medium?

The Clonetics™ and Poietics™ Basal Media are primarily composed of trace metals, amino acids, vitamins, salts, and water. The basal medium contains no growth factors or antibiotics and is not sufficient to support cell attachment and growth. To the...

What is in your ReagentPack™ Subculturing Reagents Kit (CC-5034)?

The ReagentPack™ Subculturing Reagents Kit is composed of three components: - 100 ml of HEPES Buffered Saline Solution, - 100 ml of Trypsin/EDTA- 100 ml of Trypsin Neutralizing Solution (TNS).

Can you use the AdipoRed™ Assay on cells other than preadipocytes?

Yes, the AdipoRed™ Assay Reagent can be used on many other cell types that accumulate triglycerides. It may have to be optimized for specific applications depending on the lipid that is stained.

Do I need to heat inactivate the FBS contained in the SingleQuots™ Kit?

No, we do not recommend heat inactivating the FBS provided in the SingleQuots™ Kit. Heat inactivating the serum can alter cell performance and denature essential components in the serum.
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