Data Type
Cell Information
(842)
Citations
(4649)
FAQ
(673)
Culture Media
(93)
Category
Primary Cells and Media
(265)
Transfection
(172)
Laboratory Instrumentation
(64)
Electrophoreses and Analysis
(54)
Bioassay
(48)
3D Cell Culture
(42)
Live Cell Imaging
(34)
Mycoplasma Detection and Prevention
(29)
Endotoxin Detection
(27)
Serum-free and Speciality Media
(11)
Classical Media and Reagents
(8)
Uncategorized
(7)
Cell Lines and Primary Cancer Cells
(5)
+ Show All
Research Area
Uncategorized
(172)
Basic Research
(145)
Cancer Research/Cell Biology
(81)
Immunotherapy / Hematology
(66)
Stem Cells
(54)
Molecular Biology
(39)
Neurobiology
(38)
Respiratory Research
(31)
Toxicology
(28)
Endotoxin Testing
(27)
Cardiovascular
(26)
Regenerative medicine
(23)
Dermatology/Tissue Engineering
(21)
Gene Expression
(13)
Gastroenterology
(4)
Parasitology
(3)
+ Show All
673 results sorted by
relevance
alphabetical
newest first
oldest first
G
e
n
e
r
a
t
i
o
n
o
f
a
s
t
a
b
l
e
c
e
l
l
l
i
n
e
:
A
p
o
s
i
t
i
v
e
c
o
n
t
r
o
l
p
l
a
s
m
i
d
w
o
r
k
s
,
b
u
t
I
c
a
n
n
o
t
o
b
t
a
i
n
s
t
a
b
l
e
c
l
o
n
e
s
w
i
t
h
m
y
c
o
n
s
t
r
u
c
t
o
f
i
n
t
e
r
e
s
t
.
W
h
a
t
c
o
u
l
d
b
e
t
h
e
r
e
a
s
o
n
?
One reason could be that your gene product is toxic to the cells. We recommend testing the construct for expression of the gene of interest by transient Nucleofection™ of your cells. If the proportion of expressing cells drops between 4 and 48 hours...
H
o
w
l
o
n
g
d
o
e
s
i
t
t
a
k
e
t
o
o
b
t
a
i
n
s
t
a
b
l
e
t
r
a
n
s
f
e
c
t
a
n
t
s
?
Depending on individual cell type and doubling rate, selection of stable transfectants will take between 7 and 28 days. Expansion and characterization of single cell clones will take several weeks in addition.
I
h
a
v
e
a
v
e
r
y
h
i
g
h
t
r
a
n
s
f
e
c
t
i
o
n
e
f
f
i
c
i
e
n
c
y
b
u
t
m
o
s
t
o
f
m
y
c
e
l
l
s
d
i
e
d
u
r
i
n
g
s
e
l
e
c
t
i
o
n
,
i
s
t
h
i
s
t
o
b
e
e
x
p
e
c
t
e
d
?
This is the normal pattern you should expect to see. Only a small proportion (1/10,000 to 1/100) of all transfected cells will integrate the transfected DNA into their genome and become stable transfectants. The remaining cells lose the transfected...
W
h
a
t
s
e
l
e
c
t
i
o
n
m
a
r
k
e
r
s
c
a
n
I
u
s
e
f
o
r
s
t
a
b
l
e
t
r
a
n
s
f
e
c
t
i
o
n
s
?
The most commonly used marker is the neomycin phosphotransferase (neo) gene that confers resistance to G418 to eukaryotic cells. Other markers are Puromycin, Hygromycin, Zeocin, or the HPRT gene that can be used in HPRT-deficient cells.
A
t
w
h
i
c
h
t
i
m
e
p
o
i
n
t
a
f
t
e
r
N
u
c
l
e
o
f
e
c
t
i
o
n
™
s
h
o
u
l
d
I
s
t
a
r
t
s
e
l
e
c
t
i
o
n
o
f
s
t
a
b
l
e
i
n
t
e
g
r
a
t
i
o
n
?
Antibiotics should be added 24-48 h after Nucleofection.
H
o
w
o
f
t
e
n
s
h
o
u
l
d
I
c
h
a
n
g
e
t
h
e
m
e
d
i
u
m
d
u
r
i
n
g
s
e
l
e
c
t
i
o
n
o
f
s
t
a
b
l
e
c
l
o
n
e
s
a
f
t
e
r
N
u
c
l
e
o
f
e
c
t
i
o
n
™
?
Every 2-3 days. This eliminates potentially toxic substances produced by dying cells and secondly, it keeps the concentration of the antibiotic at a constant level.
H
o
w
m
a
n
y
s
t
a
b
l
e
c
l
o
n
e
s
w
i
l
l
I
g
e
t
f
r
o
m
o
n
e
t
r
a
n
s
f
e
c
t
i
o
n
?
This depends on the respective cell type (transfection efficiency, viability and integration frequency). If it is important to you to know the frequency of stable integrants before carrying out your actual experiments, the frequency should be...
F
o
l
l
o
w
i
n
g
N
u
c
l
e
o
f
e
c
t
i
o
n
™
,
I
c
a
n
n
o
t
o
b
t
a
i
n
s
t
a
b
l
e
c
l
o
n
e
s
f
r
o
m
s
i
n
g
l
e
c
e
l
l
s
,
w
h
a
t
c
o
u
l
d
b
e
t
h
e
r
e
a
s
o
n
?
Some cell lines need certain cell densities to proliferate. For those cells you can try using conditioned medium for setting up limiting dilution or culturing single cells together with feeder cells.
I
n
m
y
N
u
c
l
e
o
f
e
c
t
i
o
n
™
E
x
p
e
r
i
m
e
n
t
I
s
e
e
l
o
w
e
r
e
x
p
r
e
s
s
i
o
n
w
i
t
h
m
y
I
R
E
S
c
o
n
s
t
r
u
c
t
i
n
c
o
m
p
a
r
i
s
o
n
w
i
t
h
y
o
u
p
m
a
x
-
G
F
P
c
o
n
t
r
o
l
.
D
o
y
o
u
h
a
v
e
a
n
y
i
n
f
o
r
m
a
t
i
o
n
a
b
o
u
t
t
h
e
d
i
f
f
e
r
e
n
t
e
x
p
r
e
s
s
i
o
n
p
r
o
f
i
l
e
s
?
The original attenuation of the IRES sequence was performed to allow for a greater difference between the expression levels of the upstream gene of interest and the downstream reporter gene. If the downstream reporter gene, the product of less than...
W
h
a
t
a
n
t
i
b
i
o
t
i
c
c
o
n
c
e
n
t
r
a
t
i
o
n
s
h
o
u
l
d
I
u
s
e
f
o
r
s
e
l
e
c
t
i
n
g
s
t
a
b
l
e
t
r
a
n
s
f
e
c
t
a
n
t
s
?
Please click on the associated file.
PREVIOUS
PAGE
5
PAGE
6
PAGE
7
PAGE
8
PAGE
9
PAGE
10
PAGE
11
PAGE
12
PAGE
13
NEXT