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306 results sorted by
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How long is the MycoAlert® Mycoplasma Detection Assay signal
stable
?
Please follow the protocol exactly as written – add reagent, wait 5 min and read, add substrate, wait 10 minutes and read again.
Missing:
cell
lines
When nucleofecting cancer
cell
lines
, do I need to be concerned about passage number?
For the most efficient gene transfer, we recommend using
cells
that are in logarithmic growth phase and at a passage number of 10-15 (from the time of thaw). This is because some
cell
lines
differentiate and change their features after many passages.
Missing:
stable
What antibiotic concentration should I use for selecting
stable
transfectants?
We highly recommend to perform a titration for G418 in a range of 0,1 mg/ml to 1,5 mg/ml; in serum-free culture expand range down to 20 µg/ml).Choose the concentration which is 0,1 or 0,2 mg/ml above the one which shows complete
cell
death...
Missing:
lines
Do you have any tips for the culture of primary
cell
s or
cell
line prior to Nucleofection?
Yes. Please see our Techincal Reference Guide which contains many helpful tips.You can find additional information in the Optimized Nucleofection Protocol for your
cell
type of interest.
Missing:
stable
At which time point after Nucleofection® should I start selection of
stable
integration?
Antibiotics for selection should be added 24-48 h after Nucleofection®.
Missing:
cell
lines
How often should I change the medium during selection of
stable
clones after Nucleofection®?
The culture medium should be changed every 2-3 days during the selection process.This eliminates potentially toxic substances produced by dying
cells
and secondly, it keeps the concentration of the antibiotic at a constant level.
Missing:
lines
What are the critical steps for successful Nucleofection® of monocytic
cell
lines
like THP-1, HL60 and U-937?
To achieve optimal results, we strongly recommend following the Optimized Protocols in regard to
cell
culture conditions (medium, splitting cycle, density before Nucleofection), DNA amount and purity.Please also be sure not to exceed 90xg when...
Missing:
stable
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
...
Missing:
stable
What should I pay attention to when transferring my reporter gene set up from a
cell
line to a primary
cell
?
In general the overall expression level differs between different
cell
types. Don't expect the same level of gene induction when working with primary
cells
. You might need to change to other DNA amounts or ratios.
Missing:
stable
I am using the acute leukemia HL-60
cell
line for CAR-T
cell
s experiments. Do you have a transfection protocol for genome editing?
The HL-60
cells
are often used as a model for in vitro research for treatment of AML (acute myeloid leukemia) in combination with modified primary human CAR- T
cells
(T
cells
expressing chimeric antigen receptors).Transfection protocols can be found...
Missing:
stable
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