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Do I need to shake the plate after adding the
Cell
Lysis Reagent during the ViaLight™ Assay?
No, this is not recommended.Shaking the plate during this step can - cause frothing which will deflect the light signal away from the detection unit, - reducing the number of RLUs observed - and producing an artificially low result.
Missing:
stable
lines
Can I take the ToxiLight™ Non-Destructive Cytotoxicity BioAssay Kit to look at the kinetics of
cell
death?
Yes, by taking a sample of the culture media (usually 20µl) into a fresh luminometer plate this can be assayed with the Adenylate Kinase Detection Reagent supplied in the Toxilight kit to look for increased levels of AK over the control. The rest of...
Missing:
stable
lines
Where are Lonza’s Normal Human Neural Progenitor (NHNP) and Normal Human Astrocytes (NHA)
cell
s isolated from?
Lonza’s Normal Human Neural Progenitor (NHNP) and Normal Human Astrocytes (NHA)
cells
are primarily isolated from the cerebral cortex (or forebrain from cerebral hemisphere) from fragmented fetal brain tissue (16-20 weeks old).
Missing:
stable
lines
When I transfected my
cell
s, error 8 appeared on the display of the Nucleofector® IIN, IIS or 2b. What is the reason?
...to the incomplete delivery; however, the
cells
can still be plated for analysis. This generally occurs because the conductivity of the solution was outside of accepted limits – this can happen if the media was not completely aspirated before resuspending, too much...
Missing:
stable
lines
How much sample do you need for accurate results with MycoAlert® Assay? Do you need
cell
s or just supernatant?
The standard protocol calls for taking 2 ml of culture media, spinning down any
cells
and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.
Missing:
stable
lines
Can I use the ToxiLight™ Non-Destructive Cytotoxicity BioAssay Kit to assess the effect of transfection procedures on the health of my
cell
s?
...Kinase are present in your transfected
cells
culture supernatant indicating that they have been damaged by the transfection procedure. This would impact on your transfection efficiency values...
Missing:
stable
lines
I tested my
cell
culture supernatant negative for mycoplasma contamination using the MycoAlert® Mycoplasma Detection Kit or the MycoAlert® PLUS Mycoplasma Detection Kit. But when I tested the same
cell
s using a PCR-based Assay, I obtained a positive result.
It is possible that the PCR is picking up dead mycoplasma whereas the MycoAlert® and MycoAlert® Plus Assay will pick up only viable mycoplasma. It is possible of course that there is contamination in the PCR giving a false positive.
Missing:
stable
lines
What optimization is necessary to get the Nucleofector® Technology to work?
None. For each
cell
type in our product list we offer a
cell
-type specific solution and thoroughly optimized electrical parameters. These are already pre-programmed in the Nucleofector® Device. Additionally our optimized protocols give you...
Missing:
stable
What
cell
number do I need per well when using the 96-well Shuttle® Device or the 4D-Nucleocuvette® Stripes?
The
cell
number is very much dependent on the
cell
type you use. For high throughput Nucleofection®, we usually start with one fifth of the optimal amount recommended for the standard Nucleofection® in the 100µl cuvette. Currently the absolute...
Missing:
stable
lines
In your optimized Nucleofection® Protocols for MDA-MB-231 and MDA-MB-468, you say to culture the
cell
s 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...
Missing:
stable
lines
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