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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™.

What protocol should I use for Nucleofection™ of patient derived blood samples, e.g. leukemia or lymphoma cells?

We do not have a ready-to-use protocol for Chronic lymphocytic leukemia (CLL) or other blood cell derived cancer cells. The cells often have a chromosomal aberration and show the properties of a cell line. Therefore we suggest to perform an...

Is it possible to transfect siRNA into blood cells from NHL patients?

Yes. Twin et al ( Blood 2007 Sep 1;110(5):1631-8.) optimized the conditions with pmax-GFP DNA and got 50-75% transfection efficiency for the patient derived diffuse large B-cell lymphoma (DLBL) and mantle cell lymphoma (MCL) cells. This set up was...

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...

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 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.

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...

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...

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...

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