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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; final...

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.

Component B in my Mouse T cell kit became solid at 4°C . Is this normal?

Yes, it is normal for component B to become solid at 4°C. The crystals dissolve at room temperature and functionality of component B is not affected.

Can I open the 96-well Shuttle™ software files by double-clicking on the file name?

No. Files can only be opened via the software.

Do you have data about the in vivo implantation of hepatocytes transfected by Nucleofection™? For how long could protein expression be detected?

There is a publication reporting on the successful reinjection of modified hepatocytes into mice.Chen et al. were able to monitor the expression over several days. This group worked with a preliminary test solution for mouse hepatocyte as the...

Are rat and mouse hepatocytes transfected by Nucleofection™ still functional?

Yes, transfected mouse hepatocytes show albumin synthesis rates comparable to those of untransfected cells. Moreover the morphology and polarization of rat hepatocytes is not affected by Nucleofection™.

Which reporter gene should I choose for my reporter gene experiment?

In general, all reporter genes can be used. For some applications (e.g. reporter gene experiments in various primary cells, experiments with late analysis time point due to starving and /or stimulation) a reporter gene with a longer half life than...

What is the maximum amount of DNA I can use in my Nucleofection™ Experiment?

The overall amount of 5 µg should not be exceeded. Look in the cell specific Optimized Protocol (OP) for the recommended total DNA amount of the cell type of interest.

When should I begin looking for the Luciferase signal after Nucleofection™?

The time course of plasmid expression after Nucleofection™ might be different from the time course seen with other transfection methods. We recommend looking at different time points and start with the first time point as early as 4 to 6 hours post...

Generation of a stable cell line: A positive control plasmid works, but I cannot obtain stable clones with my construct of interest. What could be the reason?

One reason could be that your gene product is toxic to the cells. We recommend testing the construct for expression of the gene of interest by transient Nucleofection™ of your cells. If the proportion of expressing cells drops between 4 and 48 hours...

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