0.5-1 x 105 1.5 50 0.15
24 well 0.5-1 x 105 2-5 x 105 6 100 0.5
12 well 1-2 x 105 2.5-10 x 105 12 100 1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x
0.5-1 x 105 1.5 50 0.15
24 well 0.5-1 x 105 2-5 x 105 6 100 0.5
12 well 1-2 x 105 2.5-10 x 105 12 100 1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x
viability is at least
90% and the cells are in the mid-logarithmic growth
phase. Cells grown in serum-containing medium should
be inoculated at a viable cell density of 2 x 105 cells/ml
in a 1:1 mixture
0.5-1 x 105 1.5 50 0.15
24 well 0.5-1 x 105 2-5 x 105 6 100 0.5
12 well 1-2 x 105 2.5-10 x 105 12 100 1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x
0.5-1 x 105 1.5 50 0.15
24 well 0.5-1 x 105 2-5 x 105 6 100 0.5
12 well 1-2 x 105 2.5-10 x 105 12 100 1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x
that cell
viability be >90% and the cells are in mid-logarithmic
growth phase. Cells grown in serum-containing medium
should be inoculated at a viable cell density of >3 x 105
cells/ml in a 1:1 mixture
cells from serum-supplemented medium
to EX-CELLTM Sp2/0 supplemented with 8 mM L-glutamine
at a minimum seeding density of 3 x 105 cells/mL in shaker
flasks.
2. Incubate the flasks at 37 C in a humidified
by planting new cultures
at 4 x 105 cells/mL in 20 - 30 mL of EX-CELLTM CD CHO
Fusion.
2. Subculture stocks every three to four days at a seeding
density of 4 x 105 cells/mL.
3. Continue stocks
by planting new cultures at
4 x 105 cells/mL in 20 - 30 mL of EX-CELL CD CHO Fusion.
2. Subculture stocks every three to four days at a seeding
density of 4 x 105 cells/mL.
3. Continue
by planting new cultures
at 4 x 105 cells/mL in 20 - 30 mL of EX-CELLTM CD CHO
Fusion.
2. Subculture stocks every three to four days at a seeding
density of 4 x 105 cells/mL.
3. Continue stocks
disturbing the cell pellet.
3. Resuspend the cells in EX-CELLTM 293 medium at a density
of 6 x 105 cells/mL in shaker flasks.
4. Allow the cells to adapt to EX-CELLTM 293 for an additional
4 - 6
cells were seeded at 4 x 105 cells/mL in 125 mL
shaker fl asks containing 35 mL of EX-CELL 293. Flasks
were incubated in a 10% CO2 atmosphere at 37 C, with a
shaker speed of 90 - 100 rpm
1. Subculture the cells from serum-free medium directly into
EX-CELL™ VPRO at a density of 2.5-3 x 105 cells/mL.
2. Allow the cells to adapt to EX-CELL™ VPRO Medium for
an additional 4 - 6 passages.
. Culture the cells to a density of 3-5 x 105 cells/mL in
EX-CELLTM 610-HSF medium containing 5% gamma
irradiated Fetal Bovine Serum (FBS) (Catalog No. 12107C).
Maintain the cells through 2 passages at
serum.
1. Choose cultures in logarithmic growth with viabilities above
90%.
2. Prepare a freezing medium consisting of 90% cold
EX-CELLTM MDCK medium and 10% dimethyl sulfoxide
(DMSO).
3. Using
that should be plated at Day 0 in order to have the cells reach 90% confluence by day 3. Plate out
a range of cell numbers from 1 x 105 to 1 x 106 cells per well of a
that should be plated at Day 0 in order to have the cells reach 90% confluence by day 3. Plate out
a range of cell numbers from 1 x 105 to 1 x 106 cells per well of a
viability is at least
90% and the cells are in the mid-logarithmic growth
phase. Cells grown in serum-containing medium should
be inoculated at a viable cell density of 2 x 105 cells/mL
in a 1:1 mixture
test is assessed by using BHK21
suspension cell line. Cells are seeded at a density
of 3 × 105 cells/mL and cultivated for 72 hours in
shaker flasks in standardized cell
of sin-
gle copy genes:
1 �g human genomic DNA = 3 × 105 target molecules
10 ng yeast DNA = 3 × 105 target molecules
1 ng E. coli DNA = 3 × 105 target molecules
Component Volume Concentration
viability is at least
90% and the cells are in the mid-logarithmic growth
phase. Cells grown in serum-containing medium should
be inoculated at a viable cell density of 2 x 105 cells/ml
in a 1:1 mixture
HEK293 Viral Vector Medium?
We recommend inoculating cultures at
5.0 105 viable cells/mL for 3-day cultures or
3.0 105 viable cells/mL for 4-day cultures.
6. What is the suggested cell
Biol.,
74(3), 313-330 (1973).
8. Shiozaki, K., and Yanagida, M., Mol. Cell. Biol.,
11(12), 6093-6102 (1991).
9. Xiang, H. et al., Amino Acids, 27(1), 101-105
(2004).
10. Rajesh, M. et al.
CD61. The apparent
molecular weight of the GPIIIa by SDS-PAGE in immunoprecipitaton is 105 kDa reduced and
90 kDa unreduced (prelabelled surface proteins). Ligands are fibronectin, fibrinogen, von
Willebrand
-181 (1989).
6. Brown, D. R., Trends Neurosci., 24, 85-90 (2001).
7. Kretzschmar, H. A. et al., Arch. Virol. Suppl., 16,
239-249 (2000).
8. Rezaie, P. and Lantos, P. L., Brain Res. Rev., 35
the requirements Pass
Specific rotation, [α]D25
5 mg/ml solution in alcohol
between +105° and +112° +108°
Assay high pressure
liquid chromatography
not less than 97.0% and not