Broth Agar 3.38 x 106 4.2 x 106 3.30 x 106 6.5 x 106 a
Potato Dextrose Agar 1.93 x 106 2.1 x 106 3.56 x 106 1.31 x 107 a
Sodium Azide
Tryptic Soy Broth Agar
new 1.5 mL tube, or to several tubes in conveniently-sized aliquots for storage. Chromatin from 10 9. × 106
cells (˜100 µL) will be used per ChIP for a rare transcription factor target. See Table 1 for recommended
∼150,000)
nmoles/ml = mg protein/ml × 106
mw of IgG
(106 is conversion from moles/L to nmoles/ml)
nmoles/ml = 4.5 mg/ml × 106
150,000
nmoles/ml = 30
3. To determine the number of nmoles
different
cell densities from 10 x 106 VC/mL (control) up to 100 x 106
VC/mL. Vials were thawed in a water bath at room temperature as
previously described, used for inoculation of 30 mL bioreactor
tubes, and growth
the culture is stable with a viability of > 95% and a cell density
above 1.5×106 viable cells/mL
- Prepare a MCB (min. 30 tubes in Cellvento® BHK-200 medium +
10% DMSO, 1×107 viable cells/mL)
Passage when VCD is
above 1.5 × 106 cells/mL
• When the culture is stable with a viability of > 95%
and a cell density above 1.5 × 106 cells/mL.
• Prepare a MCB (min. 30 tubes in
Gene Therapy Medium-2. Add 9 ml of fresh
Gene Therapy Medium-2.
7. Transfer the cell suspension to a T-75 flask
containing fresh Gene Therapy Medium-2 at a final
volume of 30 ml.
Adaptation to
at room temperature.
9. Aspirate supernatant and resuspend pellet in 500 l of citrate buffer solution.
10. Using citrate buffer solution, adjust the cell concentration to 2.5 x 106 cells/ml.
IV. Paraffin-embedded
Z40,627-9 Z40,629-5
1000mL Z40,628-7 Z40,630-9
Transfer needles
12 gauge SS, double-ended with one deflecting tip and one flat-cut end. For
fabrication of transfer lines with CHEM-FLEX™ 106
tubing.
µl of diluent. Incu-
bate at room temperature (18 - 22 °C) for 30 min-
utes. Protect from light at this and all subsequent
steps.
9. a. If whole blood is used, use lysing solution after
incubation
allow cells to reach viable cell
density (VCD) greater than 8 106 viable cells/mL
for optimal performance. Discard cells after
30 passages.
3. Subculture cells for at least three passages prior
1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x 105 2.5-5 x 106 60 400 4.0
90-100 mm
dish 15-30 x 10
5 5-10 x 106 150 800 8.0
T-25 flask 5-10 x
1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x 105 2.5-5 x 106 60 400 4.0
90-100 mm
dish 15-30 x 10
5 5-10 x 106 150 800 8.0
T-25 flask 5-10 x
1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x 105 2.5-5 x 106 60 400 4.0
90-100 mm
dish 15-30 x 10
5 5-10 x 106 150 800 8.0
T-25 flask 5-10 x
.
Product Profile
When assayed by flow cytometric analysis, using 10 µl
of the antibody per 1 X 106 cells and ExtrAvidin -FITC
Conjugate (Product No. E 2761), a fluorescence
intensity is observed similar
1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x 105 2.5-5 x 106 60 400 4.0
90-100 mm
dish 15-30 x 10
5 5-10 x 106 150 800 8.0
T-25 flask 5-10 x
1.0
6 well 2-5 x 105 1-2 x 106 30 200 2.0
60 mm dish 5-10 x 105 2.5-5 x 106 60 400 4.0
90-100 mm
dish 15-30 x 10
5 5-10 x 106 150 800 8.0
T-25 flask 5-10 x
µl of diluent. Incu-
bate at room temperature (18 - 22 °C) for
30 minutes. Protect from light at this and all
subsequent steps.
9. a. If whole blood is used, use lysing solution after
incubation
or 1 x 106 cells per
tube.
2. Add 5 µl of monoclonal antibody to tube(s) contain-
ing cells to be stained. Vortex tube gently. Incu-
bate the cells at room temperature (18–22 °C) for
stained, i.e. 1 µg of monoclonal
antibody per 1 x 106 cells in a final volume of 100 µl.
Tap tube gently to mix. Incubate the cells on ice for
30 minutes.
Proper controls to
for 15 minutes in the dark.
9. Spin down cells at 2500 rpm (600Xg) for 3 minutes and discard buffer.
10. Resuspend the cells in 1mL of 1X Assay Buffer/5X 106 cells by gently pipetting up
for 15 minutes in the dark.
9. Spin down cells at 2500 rpm (600Xg) for 3 minutes and discard buffer.
10. Resuspend the cells in 1mL of 1X Assay Buffer/5X 106 cells by gently pipetting up