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523 results sorted by
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C
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There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present.
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S
h
u
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t
l
e
™
d
e
v
i
c
e
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In addition to proper cell handling and using culture media optimized for long term culturing of hepatocytes (Lonza Basal medium supplemented with BulletKit), we recommend separating the healthy hepatocytes from the dead cells and debris in the...
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N
u
c
l
e
o
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e
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t
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o
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™
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F
o
r
h
o
w
l
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g
c
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l
d
p
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x
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s
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n
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There is a publication reporting on the successful reinjection of modified hepatocytes into mice.Chen et al. were able to monitor the expression over several days. This group worked with a preliminary test solution for mouse hepatocyte as the...
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M
y
c
o
A
l
e
r
t
®
/
M
y
c
o
A
l
e
r
t
®
P
L
U
S
R
e
a
g
e
n
t
a
n
d
S
u
b
s
t
r
a
t
e
?
For optimal assay conditions, reconstituted reagent and substrate should be used fresh within 2 hours after reconstitution.During this time they can be kept at room temperature.For the MycoAlert® Kit: Unused, reconstituted components can be aliquoted...
A
r
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r
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s
f
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d
b
y
N
u
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l
e
o
f
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t
i
o
n
®
s
t
i
l
l
f
u
n
c
t
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o
n
a
l
?
Yes, transfected mouse hepatocytes show albumin synthesis rates comparable to those of untransfected cells. Moreover the morphology and polarization of rat hepatocytes is not affected by Nucleofection®.
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o
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®
A
s
s
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y
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D
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n
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?
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.
W
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H
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m
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H
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p
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p
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r
t
o
N
u
c
l
e
o
f
e
c
t
i
o
n
™
?
In order to avoid cell compaction in the pellet and difficulties in cell resuspension, we recommend using round bottom 2 ml vials or 50 ml BD Falcon™ tubes for all centrifugation steps prior to Nucleofection™.
I
s
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q
u
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t
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f
y
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w
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M
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c
o
A
l
e
r
t
®
M
y
c
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p
l
a
s
m
a
D
e
t
e
c
t
i
o
n
A
s
s
a
y
?
No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.
W
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H
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n
H
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p
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t
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w
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t
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t
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e
N
u
c
l
e
o
f
e
c
t
o
r
®
?
We recommend allowing the transfected hepatocytes to recover in Plating medium for 72 hours post-Nucleofection®.For induction studies (e.g. CYP3A4), the Plating medium is replaced with maintenance medium (William's E without FCS and EGF) and...
W
h
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g
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M
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c
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®
M
y
c
o
p
l
a
s
m
a
D
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t
e
c
t
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o
n
A
s
s
a
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,
w
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i
s
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t
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s
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t
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s
p
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c
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?
It is highly recommended that the samples are spun down prior to testing. If cells are present in the assay they will contribute to the initial reading producing a higher background. This in turn can decrease the difference between the background and...
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