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I observed high mortality rates in mES cells after Nucleofection with a Cre recombinase expression vector. How can I increase the viability?

The High mortality after transfection might be due to the Cre recombinase itself, because the mammalian genome contains pseudo loxP sites (Thyagarajan B et al. (2000) Mammalian genomes contain active recombinase recognition sites. Gene....

Can Lonza's antibiotics be used in clinical trials?

No. They are for research purposes only.

What is the basic principle of the Nucleofector Technology?

The Nucleofector™ Technology is a novel transfection method especially designed for the needs of primary cells and difficult-to-transfect cell lines. It is a non-viral method based on a unique combination of electrical parameters and cell-type...

Why is the Nucleofector Technology ideal for primary cells and difficult-to-transfect cell lines?

There are several reasons to choose the Nucleofector™ Technology for your gene transfer experiments. One is the direct transfer of DNA to the cell nucleus. This makes gene expression independent from cell division. Therefore, the technique is the...

Do you have a protocol for stimulation of mouse B cells?

Yes, if you want to transfect mouse B cells, we recommend to stimulate the cells before. The information is also available in the mouse B cell nucleofection protocol. Our R&D has used following combination : RPMI1640 [Lonza; Cat. No....

What is the typical cell distribution for peripheral blood mononuclear cells (PBMC) catalog number CC-2702? 

We don't characterize each lot of these cells. Therefore, we don't make any guarantee regarding cell type distribution.Typically, you would expect to see approximately 60-80% T Cells, 10% monocytes, and the remaining cells would be Beta cells,...

Which mobilization regime has been used for the CD34+ cells from mobilized peripheral blood (4Y-101)? Can I also have the cells from donors treated with another mobilization regime?

The mobilized peripheral blood used for 4Y-101 is derived  from peripheral blood mobilized with Neupogen® 10 mcg/kg/day x 5 days. The cryopreserved cells can be isolated from day 5 and/or day 6 collections. It is possible to request...

Does DMSO (ex. brought in with the substrate) affect Nucleofection?

Most research paper agrees that the upper limit of cell tolerance to DMSO presence in their cell culture media is about 1% final DMSO (this might vary with the cells and media).In the case of the Nucleofection, the DMSO will likely be transfected (at...

Can I replace the RAFT collagen with another hydrogel type ex. Matrigel?

Complete substitution for collagen type I within the RAFT system (including its absorption step) is not possible, but one can certainly add Matrigel or laminins to the RAFT collagen mixture, up to a certain ratio. The absorption conditions (ex. time)...

What is the Cocoon® technology? 

The Cocoon® Platform utilizes a single use, disposable cassette to functionally close and automate cell manufacturing. Primarily developed to automated all steps in CAR-T cell manufacturing, the Cocoon® Platform is being used for numerous...
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