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How many cortical neurons can I expect from a single pup?

According to QBM Cell Sciences, you can expect 12 million cortical neurons from an average E18-E19 rat pup. You can expect 4 million cortical neurons from an average mouse pup.

Does the 96-well Shuttle recognize if the cuvette plate is in backwards?

No, it does not recognize the orientation of the plate. Please make sure that well A1 is positioned in the upper left corner of the retainer.

Do any of the three instruments used in the 96-well Shuttle System "go to sleep" or power off automatically?

The energy options of the laptop are set as such that only the screensaver is activated, the harddisk won't turn down and hibernation is disabled. One can change these settings via the system control. However this is not recommended because by...

What is the molecular weight of your control pmaxGFP plasmid?

The molecular weight is approximately 2.3x10e6 g/mol based on the average MW of a base pair being 660 g/mol.

In my Nucleofection Experiment I see lower expression with my IRES construct in comparison with you pmax-GFP control. Do you have any information about the different expression profiles?

The original attenuation of the IRES sequence was performed to allow for a greater difference between the expression levels of the upstream gene of interest and the downstream reporter gene. If the downstream reporter gene, the product of less than...

What are the best time points for analysis of my siRNA experiments?

As the stability and half-life of various mRNAs and their protein products varies, it is important to empirically determine the best time points for assessing target knockdown. For example, it has been documented that in mammalian cells, mRNA...

Can I co-transfect oligonucleotides and plasmids with the Nucleofector Technology?

Yes. As the same protocol applies for any nucleic acid substrate (vectors or oligonucleotides) you can easily perform co-transfections either for transfection control or rescue experiments.

I want to stimulate human primary T-cells with anti-CD3 and anti-CD28. Do you have a detailed protocol for this?

For optimal stimulation of the transfected cells we recommend coating 96-wells plates for stimulation [Nunc Immuno™ Plate C96 Maxi Sorp™, Cat. No. 430 341] with 50 µl of a solution of anti- CD3 antibody [OKt 3; eBioscience, Cat. No. 14-0037-82;...

Why does Lonza recommend using non-ventilated caps when culturing insect cells?

The reason is that insect cells are often incubated in very simple incubators that are not humidified (and without CO 2 at 24°C to 28°C). Insect media typically do not need CO 2 for buffering pH, so gas exchange is not required. To avoid...