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Do I need to use a gelatin, fibronectin, collagen, or Matrigel™ coating when culturing my Clonetics™
Cell
s?
Most Clonetics™
Cells
do not require an extracellular matrix when grown on tissue culture treated plastic – flasks, dishes, well plates (with the exception of hepatocytes, and the rat/mouse neuronal and dorsal root ganglion neurons, rat cardiac...
Missing:
stable
lines
Does it matter if the PBS used for monocyte enrichment contains calcium and magnesium, if the
cell
s should be used later in Nucleofection®?
The PBS should be calcium and magnesium free to prevent clumping of
cells
. We routinely use PBS without calcium and magnesium for example Lonza Cat. No. BEBP17-516Q
Missing:
stable
lines
Why do you recommend 7AAD instead of PI staining for
cell
determination in mouse macrophages?
The results obtained using 7AAD staining are much clearer than those obtained using propidium iodide (PI). Using flow cytrometry stained and unstained, transfected and non-transfected populations can be separated much more easily.
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
Are the human mesenchymal stem
cell
s (PT-2501) more prone to become differentiated down one lineage over the other?
...differentiation if allowed to reach confluence. As such, to maintain
cells
as undifferentiated
cells
, it is critical to subculture
cells
when they are subconfluent....
Missing:
stable
lines
How many blood donations are pooled for pMAT
cell
s included in the Pyro
Cell
® Monocyte Activation Test (MAT) Kit?
One production lot of pMAT
cells
contains equal amounts of peripheral blood mononuclear
cells
from four blood donations.
Missing:
stable
lines
Do I need to shake the plate after adding the
Cell
Lysis Reagent during the ViaLight™ Plus 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
Should I add any nutritional supplementation to TheraPEAK 293-GT® Media System to support
cell
growth?
It is not necessary to add nutritional supplementation because TheraPEAK 293-GT® Media System is preformulated with 4 mM GlutaMAXTM, however if your process uses an alternative nutritional source you can add any supplementation required.
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
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
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