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How many cells do I have to use for each well of the 16-well Nucleocuvette® Strip?

This depends on the cell type you are working with. In our Optimized Protocols the cell numbers range from 2.5 x 10^4 cells (mouse DC) up to 1x 10^6 cells (human T cells). Detailed information is provided in our 4D-Nucleofector® Optimized Protocols.

Can I use SDS as a detergent for a positive control if I do cytotoxicity assay with ToxiLight?

No. SDS has a negative influence on Adenylate Kinase (AK) activity. SDS is a cationic detergent, it is a very powerful protein denaturing agent, you will be aware of its use in SDS-PAGE for those reasons. ToxiLight™ works by measuring the activity of...

Do I have to apply the same program to each well of the 16-well Nucleocuvette®?

No, each well can be addressed separately. You may apply up to 16 different programs to one 16-well Nucleocuvette® Strip.

How many 100 µl Nucleocuvettes® can be transfected in parallel using the 4D-Nucleofector® System?

The 4D-Nucleofector® X Unit can work with two 100µl Nucleocuvettes® in parallel. However, the operation software allows defining up to 50 Nucleocuvettes® per experiment. These cuvettes will be transfected by the system continuously in...

Which luminometer is suited for performing the MycoAlert® Assay?

For performing the MycoAlert® Assay in theory every luminometer is suitable as long as a specific sensitivity is given. Please find attached our list with luminometers we know to be suited to run the MycoAlert® and MycoAlert® Plus Mycoplasma...

Are there positive controls for the MycoAlert® Assay?

The MycoAlert® Assay Control Set (LT07-518) is available separately and includes a lyophilized positive control (1 ml) and assay buffer (2 ml) for reconstitution. The assay buffer also serves as a negative control. The positive and negative controls...

Can I optimize the transfection efficiency or cell viability after the 4D-Nucleofector® Cell Line Optimization any further?

We offer a program fine tuning matrix for Optimization of Nucleofection® Conditions - A Short Guideline (lonza.com)Based on your current most satisfactory pulse(s) it provides guidelines which further programs to test to either increase cell...

Does the MycoAlert® Assay recognize the whole spectrum of mycoplasma?

The mycoplasma specific metabolism detected by the MycoAlert® Assay is present in all members of the mollicute family (Mycoplasma, Acholeplasma, Entomoplasma and Spiroplasma) except Ureaplasma (which are no usual suspects in cell culture). 95% of all...

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).

Do bacteria lead to false positive MycoAlert® Results?

NO - the lysis reagent in the MycoAlert® Assay is not strong enough to lyse bacteria / yeast. Also, the enzymes utilized are not very common in bacteria or yeast – but are universal among the mollicute family. Bacterial contamination is...
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