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What is the ratio of Supplement to Nucleofector® Solution?

The ratio is 1:4,5 (500 µL of supplement is added to 2.25 mL of Nucleofector® Solution).For a single 100 µL reaction, use 18 µL of supplement plus 82 µL of solution to make the 100 µL.

What is the molecular weight (MW) of your control plasmid pmaxGFP?

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


In your optimized Nucleofection® Protocols for MDA-MB-231 and MDA-MB-468, you say to culture the cells without CO2. Why?

The media recommended for these cells is Leibovitz's L-15 medium which does not contain sodium bicarbonate to act as a buffer for the carbonic acid that would normally form in the presence of CO2. Furthermore, if the cells are cultured in Leibovitz's...

Do you have any siRNA Nucleofection® results using concentrations lower than 50nM?

Yes. Please click on the attached Technical Reference Guide: "Designing an RNAi Experiment Using Nucleofection®". On page 3 you can find a table with some examples.

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

Yes, the detailed protocol is part of our 4D-Nucleofector® Protocol for stimulated Human T Cells.For optimal stimulation of the transfected cells we recommend coating 96-wells plates for stimulation [Nunc Immuno™ Plate C96 Maxi Sorp™, Cat. No. 430...

What are useful methods to enrich/purify specific hematopoetic cell populations before Nucleofection?

Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).

For hepatocytes, is plating density a concern post Nucleofection®?

Yes, the optimized protocols for the standard Nucleofector® recommend the use of 6 well plates. Plating into a 12-well plate will pose a problem as less cells will attach at high densities.

Do you recommend a purification method to purify specific blood/immune cell populations before Nucleofection®?

We recommend using negative selection (depletion) because your purified cells are "untouched", not influenced, and strictly not activated.Positive selection may cause increasing amounts of dead cells and could also lead to activation of the cells due...

Four hours after Nucleofection®, I can see the hepatocytes have attached to the well, but the morphology does not look correct, should I be concerned?

There is no reason for alarm. The hepatocytes may not exhibit normal morphology a few hours after Nucleofection® but by 24 hours post Nucleofection®, morphology should be normal. Remember to perform a fluid change about 4 hours post...
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