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Where can I find what the error codes on the Nucleofector or 96-well Shuttle mean?

In the back of the respective manuals, there is a chart giving a brief description of the error codes. If more information is needed or an error code is recurrent, it is best to contact Scientific Support for advice to determine if there is a...

According to which standard is the Nucleofector device certified?

The Nucleofector II was tested by TÜV Rheinland product safety GmbH and found to be in compliance with the following standards: IEC 61010-1:2001 (2nd edition) and EN 61010-1:2001 (2nd edition). TÜV Rheinland of North America Inc. tested the...

Is it typical for the fluorescence observed from my tagged protein to appear less than the empty fluorescent vector?

Yes, this is a routine observation. Fluorescent fusion proteins can appear less bright than the fluorescent protein itself. This may be due to differences in protein folding of the fused protein.

Why does Lonza recommend using non-ventilated caps when culturing insect cells?

The reason is that insect cells are often incubated in very simple incubators that are not humidified (and without CO2 at 24°C to 28°C). Insect media typically do not need CO2 for buffering pH, so gas exchange is not required. To avoid evaporation of...

How critical is the time point for analysis post transfection?

It is very critical. Waiting too long for analysis post transfection can lead to loss of peak expression. The optimal time for analysis is related to two factors: stability of protein being expressed and transfection method. For a short-lived...

What is the ratio of Supplement to Nucleofector solution?

The ratio is 1:4.5 (500µL's of Supplement is added to 2.25mL's of Nucleofector solution). For a single reaction, use 18 ul of supplement plus 82 ul of solution to make the 100 ul.

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...

Do we sell the Nucleofector Solution (or any other component of the Nucleofector Kit) separately?

We don't sell the individual components of the kits separately because our product line is a kit concept line to ensure the use of Amaxa™ certified products with quality controlled lot numbers that guarantee optimal results.

What are the key things in a Nucleofection that one can do to maximize viability?

Pay careful attention to centrifugation speeds prior to Nucleofection™; be sure not to centrifuge the cells at higher than 90xg. Any trypsinization should use the minimal amount of reagent and time necessary in order to minimize stress to the cells,...

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).