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Is it possible to pre-plate substrates when using the 96-well Shuttle® for Nucleofection®?

Yes, pre-plating of substrates like siRNA and subsequent storage at -20°C is possible.

What kind of protocol you recommend for stimulation of human T cells?

For getting the results we present in our datasheet, we recommend the follwing strategy: Cells should be stimulated and cultivated for 3 days before Nucleofection, preferentially on plates coated with 1 µg/ml anti-CD3 and 2 µg/ml anti-CD28 antibody....

Are there any effects on the differentiation of neural stem cells following Nucleofection®?

The ability of transfected rat or mouse neural stem cells to be subsequently differentiated into a variety of cell types is well documented in:Olig2 Overexpression Induces the In Vitro Differentiation of Neural Stem Cells into Mature...

Can calcium influx issues affect viability or differentiation in Stem Cells after Nucleofection®?

When cells are transfected by Nucleofection®, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection® in medias that contain...

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...

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...

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 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...

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...

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