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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...

Is there a reason why in our Nucleofector Protocols for Primary Human Chondrocytes we include FCS in the pronase and collagenase solutions for preparation of chondrocytes?

Yes, even though this may sound odd since serum is generally known to inactivte proteases. We recommend it in this case, however, because the incubation times are quite long and the cells are stressed by the procedure. The amounts of enzymes should...

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.

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

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

What is the molecular weight of the GFP protein expressed by your control pmaxGFP™ plasmid?

The molecular weight of the protein is about 27kDa.

What antibodies can be used against the GFP produced by pmaxGFP™ and pmaxFP™-Green vectors? What antibodies can be used to detect denatured GFP? What antibodies can be used to detect the RFP expressed by pmaxFP-Red?

Most of the commercial antibodies for GFP do not detect GFP expressed from pmaxGFP or pmaxFP-Green vectors. The origin of the protein expressed by the pmaxGFP control plasmid and pmaxFP-Green is from the green fluorescent protein CopGFP, cloned from...

What is the shelf life of the 96-well Shuttle™ kits?

The shelf life of the 96-well Shuttle™ Kits is 24 months from date of manufacturing. We guarantee a remaining shelf life of at least 6 months after sending them out of the warehouse.

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;...

Do you recommend a particular supplier for siRNA duplexes for use in Nucleofection™ ?

For siRNA applications using the Nucleofector technology we recommend siRNA duplexes from Dharmacon, part of Thermo Fischer Scientific.

Can I use the protocol for MCF7 cells for other breast cancer cell lines?

No. Often times closely related cell types will require completely different conditions for Nucleofection™. Therefore, you may want to consider performing a full optimization by using the Cell Line Optimization Kit. Please click on the related link...

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