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Do you have any recommended methods for detaching human monocytes from the plate for assaying or before Nucleofection®?
We have had good results by incubating the
cells
in ice cold PBS for 10 minutes and then rinsing the plates. Alternatively, the
cells
can be detached without a medium change by gently pipetting the
cell
suspension up and down. Our experience has...
Missing:
stable
lines
Do I need a pure neuronal culture for Nucleofection® ?
In principle, it should be absolutely fine to nucleofect pure neuron cultures (e.g. after purification by Ficoll). Glial
cells
are not necessary for transfection. What one would have to consider for example, is to get the required number of neurons...
Missing:
stable
lines
How should I purify my DNA for Nucleofection® of neurons?
...suppliers. The presence of endotoxins can increase
cell
mortality and this is especially true of sensitive
cells
such as primary neurons. The presence of endotoxins can increase
cell
mortality and this especially true of primary
cells
such as neurons...
Missing:
stable
lines
What is the difference between Lonza’s BEGM™ medium and the Growth Media portion of Lonza’s B-ALI BulletKit™ Medium?
Lonza’s BEGM™ medium and the Growth Media portion of Lonza’s B-ALI BulletKit™ Medium are essentially the same and can be used interchangeably (for example,
cells
could be cultured in Lonza’s BEGM™ medium just prior to differentiation using...
Missing:
stable
lines
What is the difference between Lonza’s SAGM™ medium and the Growth Media portion of Lonza’s S-ALI BulletKit™ Medium?
Lonza’s SAGM™ medium and the Growth Media portion of Lonza’s S-ALI BulletKit™ Medium are essentially the same and can be used interchangeably (for example,
cells
could be cultured in Lonza’s SAGM™ medium just prior to differentiation using...
Missing:
stable
lines
Can I use the Nucleofector® Technology for RNAi applications? How do I start?
Yes. The Nucleofector® Technology can be easily applied for any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the
cell
type specific Optimized Protocol for DNA vectors (cDNA, shRNA or miRNA expressing plasmids...
Missing:
stable
lines
What is the Cocoon® Technology?
The Cocoon® Platform utilizes a single use, disposable cassette to functionally close and automate
cell
manufacturing. Primarily developed to automated all steps in CAR-T
cell
manufacturing, the Cocoon® Platform is being used for numerous...
Missing:
stable
lines
How are Lonza's CD14+ monocytes isolated?
Lonza's CD14+ monocytes are isolated first by obtaining mononuclear
cells
from peripheral blood via density gradient centrifugation. The CD14+
cells
are then isolated form the mononuclear
cell
population using either positive or negative...
Missing:
stable
lines
How do you calculate a population doubling?
A population doubling is the total number of population doublings of a
cell
type since its initiation in vitro. The number of population doubling is calculated using the following equations:
Cell
count at day of passage /
Cell
count 24 hours...
Missing:
stable
lines
What is the purpose of EDTA in trypsin?
EDTA is a chelator of metal ions like calcium and magensium. It is used with trypsin because it helps bind up the divalent cations in the medium. It helps to keep the
cells
from sticking to each other. If its not there….you would need a higher...
Missing:
stable
lines
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