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106-20-7
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Process Guidance Cellvento™ BHK-200 Medium
Guidance.indd 7 03.03.14 10:28
8
Annex 1: Freezing procedure for BHK21 cells in serum-free medium
1. Materials
• Clean bench or laminar flow hood
•
The Effective Use of Protein Kinase Inhibitors
97 96 94 106 103 16 106 106 105 42 54 23
CK1
CK2 104 98 102 103 103 107 96 87 97 93 98 18 19 104 73 112 104 101 106
PHK 51 81 58
Process Guidance - Cellvento BHK-200
TPB
+10% serum
+10%
serum
+7%
serum
+5%
serum
+3%
serum
+1%
serum
+0.5%
serum
high
density
low
density
normal
culture
80
60
40
20
0
100
95
90
85
CELLINE CL 1000 OPERATING PROTOCOL WHEN USING SUSPENSION CELLS
number to about 125 x 106 and completely change the nutrient medium. Repeat
the cycle.
7-Day Feeding Schedule: Inoculate (or at steady state split the cells back to) about 25 x 106 viable cells and add
User Guide: Sf-RVN® Platform
100 x 106 cells required
If the cell density is 4.5 x 106 viable cells/mL:
7. Transfer the calculated volume of cell suspension to
one or more 15, 50, or 200 mL conical centrifuge
tubes.
8. Centrifuge
The Effective Use of Protein Kinase Inhibitors
97 96 94 106 103 16 106 106 105 42 54 23
CK1
CK2 104 98 102 103 103 107 96 87 97 93 98 18 19 104 73 112 104 101 106
PHK 51 81 58
App Note: Scalability Studies for Millistak+® HC and Millistak+® HC Pro Micro 20 Devices
details: 16.9 x 106 TC/mL, 93% viable). Filter resistance profiles are shown in the top panel and filter turbidity breakthrough curves
are shown in the lower panel. Millistak+® HC Pro Micro 20 (20 cm2) and
Growth Characteristics of the expresSFplus Serum-Free Insect Cell Line in EX-CELL420 Serum-Free Media
days then appeared
to plateau between days two to four, reaching maximum
densities of approximately 7 x 106 cells/mL. Culture viabilities
remained high (above 95%) until day four, after which
viabilities dropped
TB329 Sf9 Insect Cells
Flask Size Cell Number Medium Volume
25-cm2 flask 1.0 × 106 5 ml
75-cm2 flask 3.0 × 106 10 ml
150-cm2 flask 6.0 × 106 30 ml
Passaging Suspension Cultures
Sf9 Insect
Product Information Sheet - S6576
for up
to two weeks.
7. To pull-down the antibody-bound chromatin
complexes in ChIP, use 5 µL (for chromatin from
≤ 2 x 106 cells) or 10 μL (for chromatin from > 2 x
106 cells) of blocked
Bulletin - CHROP
that does not bind DNA directly, e.g., EZH2
qPCR 5-10 x 106
Microarray 50 x 106
(5 ChIPs)
Sequencing 100-200 x 106
(10-20 ChIPs)
Table 1: Guidelines for number of cells required, depending
Product Information Cellvento™ BHK-200 Medium
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
• Freeze some cells (WCB 7%, with 1
Data Sheet - 102376
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
• Freeze some cells (WCB 7%, with 1
Insect Directﾙ Protein Expression & Purification System
1.0 × 106/ml
20 µg + 0.5 ml 60 µl + 0.4 ml 0.5 ml 5 µl 0.25 ml 96-well filter
plate
2 × 1.5 ml 0.5 ml
100 ml 90 ml at
1.1 × 106/ml
Product Information Sheet: Cellvento® BHK-200 cell culture medium
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
• Freeze some cells (WCB 7%, with 1
Product Information Sheet - 11663925910
do not freeze. Warm to
+20 to +25°C before use.
Store 2 weeks at +2 to +8°C, or
2 days at +20 to +25°C, or 8 hours at
+30°C.
7 Washing buffer,
10x conc. (Bottle 7)
Data Sheet - G0916
by centrifugation for 5 minutes at
200 x g. Re-suspend the cells at a
concentration of 3 x 106 to 5 x 106 cells/ml in
50% fresh Gene Therapy Medium-1 and 50%
conditioned Gene Therapy Medium-1.
Supplement
Data Sheet - H6533
from the HA-
7 hybridoma produced by the fusion of mouse myeloma
cells and splenocytes from a BALB/c mouse immunized
with a synthetic peptide corresponding to amino acid
residues 98-106 (YPYDVPDYA) of
Data Sheet - H3663
antibody in PBS containing
0.05% TWEEN 20, with agitation for 120
minutes.
6. Wash the membrane three times for 5 minutes
each in PBS containing 0.05% TWEEN 20.
7. Incubate the membrane with Anti-Mouse
Data Sheet - G9916
by centrifugation for 5 minutes at
200 x g. Re-suspend the cells at a concentration of
3 x 106 to 5 x 106 cells/ml in 50% fresh Gene
Therapy Medium-3 and 50% conditioned Gene
Therapy Medium-3. Supplement
Data Sheet - I5408
TiterHigh Sf supports fast
cell growth rates (doubling times of 20-24 hours), high
cell densities of >20 x 106 cells/mL for Sf21 and >10 x
106 cells/mL for Sf9, while maintaining high cell viability
and
Critical evaluation on the use of Dmb-dipeptides in the synthesis of neurotoxic prion peptide
Ala-Gly
H-Leu-Ala
N
H
H
N
Gly-Met-Leu-OH
O
O
CH3
Fig. 7: a) HPLC profile and b) MALDI-TOF of crude PrP (106-126) prepared using 3
Dmb-dipeptide derivatives.
Novabiochem®
Critical evaluation on the use of Dmb-dipeptides in the synthesis of neurotoxic prion peptide
Ala-Gly
H-Leu-Ala
N
H
H
N
Gly-Met-Leu-OH
O
O
CH3
Fig. 7: a) HPLC profile and b) MALDI-TOF of crude PrP (106-126) prepared using 3
Dmb-dipeptide derivatives.
Novabiochem®
Microbiology - 100778 Cult-Dip combi - Instructions for use
infection
bacterial count 105–106 moderate infection
bacterial count 106 or more heavy infection
Total Bacteria Count Agar (TTC-Agar)
Bacteria
per ml 103 104 105 106 107
11-109-5_PB_100778:08
Corneal Epithelium Progenitors, Human
concentration 1 x 106 cells/mL).
5) Immediately add 1 mL cell suspension into labeled cryotubes.
6) Immediately transfer tubes to a NALGENE® Cryo 1°C freezing container (#5100-
0001).
7) Immediately
Product Information Sheet - AST1001
104
4 well plate 2 cm2 1 × 105 – 2 × 105
35 mm dish 10 cm2 5 × 105 – 1 × 106
60 mm dish 20 cm2 1 × 106 – 2 × 106
Product Information Sheet - AST1039
104
4 well plate 2 cm2 1 × 105 – 2 × 105
35 mm dish 10 cm2 5 × 105 – 1 × 106
60 mm dish 20 cm2 1 × 106 – 2 × 106
Product Information Sheet - AST1037
104
4 well plate 2 cm2 1 × 105 – 2 × 105
35 mm dish 10 cm2 5 × 105 – 1 × 106
60 mm dish 20 cm2 1 × 106 – 2 × 106
Cell Culture Media 4CHO Sample Kit
trial samples
1 0.5 x 106 1.5 x 106 3 x 106
10 5.0 x 106 15 x 106 30 x 106
15 7.5 x 106 22.5 x 106 45 x 106
30 15 x 106
Product Information Sheet- H3663
PBS containing 0.05%
TWEEN 20, with agitation for 120 minutes.
6. Wash the membrane three times for 5 minutes
each in PBS containing 0.05% TWEEN 20.
7. Incubate the membrane with
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