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H
o
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1
6
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N
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v
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t
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®
S
t
r
i
p
?
This depends on the cell type you are working with. In our Optimized Protocols the cell numbers range from 2.5 x 10^4 cells (mouse DC) up to 1x 10^6 cells (human T cells). Detailed information is provided in our 4D-Nucleofector® Optimized Protocols.
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™
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?
The GFAP expression decreases rapidly with passaging. The results that we give are performed in the 1st passage out of cryopreservation.
W
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N
u
c
l
e
o
f
e
c
t
i
o
n
™
?
Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).
D
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1
6
-
w
e
l
l
N
u
c
l
e
o
c
u
v
e
t
t
e
®
?
No, each well can be addressed separately. You may apply up to 16 different programs to one 16-well Nucleocuvette® Strip.
D
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y
o
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c
o
m
m
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n
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b
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N
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c
l
e
o
f
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c
t
i
o
n
®
?
We recommend using negative selection (depletion) because your purified cells are "untouched", not influenced, and strictly not activated.Positive selection may cause increasing amounts of dead cells and could also lead to activation of the cells due...
H
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m
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n
y
1
0
0
µ
l
N
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4
D
-
N
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f
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c
t
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®
S
y
s
t
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m
?
The 4D-Nucleofector® X Unit can work with two 100µl Nucleocuvettes® in parallel. However, the operation software allows defining up to 50 Nucleocuvettes® per experiment. These cuvettes will be transfected by the system continuously in...
W
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e
d
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d
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m
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f
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r
N
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t
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a
l
K
i
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l
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r
(
N
K
)
c
e
l
l
c
u
l
t
u
r
e
?
For the cultivation of Natural Killer (NK) cells we recommend using the X-VIVO 15 medium.In regard to cytokines, this varies depending upon the application, but many cited publications indicate using IL-2 for activation and longer term culture.
D
N
A
-
p
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i
t
y
a
n
d
N
u
c
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f
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®
:
C
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A
2
6
0
:
A
2
8
0
r
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n
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y
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d
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l
l
v
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a
b
i
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i
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y
?
Yes. To check the quality of your DNA we strongly recommend determining the A260:A280 ratio. It should be ideally 1.8 - 2.0 for a good DNA preparation, but least 1.6.
C
a
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n
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D
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N
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f
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t
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n
®
e
x
p
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r
i
m
e
n
t
s
?
Yes. You should verify the integrity of your DNA on an agarose gel to see if it is degraded. Compare undigested plasmid to plasmid DNA digested with a suitable single cut restriction enzyme to linearize. Supercoiled plasmid will run faster than...
D
o
L
o
n
z
a
'
s
N
K
c
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s
p
r
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l
i
f
e
r
a
t
e
i
n
c
u
l
t
u
r
e
?
Publications indicate that IL-2 and IL-21 can stimulate NK cell proliferation in addition to using a human serum supplemented medium.
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