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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 selection markers can I use for stable transfections?

The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.

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.

My suspension-adapted HEK293 (or CHO) cells tend to clump in culture. How can I avoid this?

Efficient protein production experiments require growth of the host cell in single-cell suspension, something that is sometimes difficult to achieve since most established cell lines retain their adherent growth characteristics. Commercially...

How can I improve Human Hepatocyte viability post Nucleofection on the 96-well Shuttle device?

In addition to proper cell handling and using culture media optimized for long term culturing of hepatocytes (Lonza Basal medium supplemented with BulletKit), we recommend separating the healthy hepatocytes from the dead cells and debris in the...

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 best way to centrifuge Human Hepatocytes prior to Nucleofection?

In order to avoid cell compaction in the pellet and difficulties in cell resuspension, we recommend using round bottom 2 ml vials or 50 ml BD Falcon™ tubes for all centrifugation steps prior to Nucleofection™.

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.