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DuoFect™
- Easy
to use - simply add the transferrin/DNA complex
directly to cells
- Highly
efficient - 30 to 1000-fold enhanced transfection
efficiency
- Flexible
- simply add complex directly to your cells in
serum- containing or serum-free medium
- Extremely
gentle - utilizes a natural biological process to
introduce DNA into the cells
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 DuoFect™
utilizes the natural endocytosis of transferrin molecules and the
associated cellular receptor to transport DNA across the cellular
membrane. Transfection rates of 100% have been shown for tumor
cell lines Neuro 2A, Jurkat and K562. In other established cell
lines, such as HeLa, CHO and COS DuoFect™ works with high
efficiencies, excellent reproducibility and with low
cytotoxicity.
The
DuoFect™ transfection system is a high-efficiency nucleic
acid delivery method based on transferrin receptor-mediated
endocytosis to carry DNA into cells. Human transferrin is
covalently linked to a polycationic carrier (polyethylenimine)
with intrinsic endosomal activity. The modified polyethylenimine
(PEI) conjugated transferrin molecules maintain their ability to
bind to their cognate receptor and to mediate efficient iron
transport into the cell. The transferrin/condensed DNA complexes
are transported across the cell membrane via endocytosis of the
DNA complex and transferrin receptor. PEI then acts as a proton
sponge resulting in endosome lysis and release of the DNA into
the cell (see figure).
Components
DuoFect conjugate (PEI-transferrin), lyophilized.
Deferrioxamine, lyophilized.
HBS buffer concentrate.
Storage
-20C
General Considerations
Quality of DNA preparation: The DNA quality is a critical
parameter for transfection. The transfected DNA should be
purified by cesium chloride gradient or commercial purification
columns and should be free from RNA contamination.
References
Kircheis, R. et al (1997). Gene Therapy, 4, 409-418.
Boussif, O. et al (1995). Proc. Natl. Acad. Sci. USA, 92,
7297-7301.
Cotten, M. et al (1992). Methodes Enzymol., 217, 618-644.
Curiel, D. T. et al (1991). Proc. Natl. Acad. Sci. USA, 88,
8850-8854.
Wagner, E. et al (1991). Proc. Natl. Acad. Sci. USA, 88,
4255-4259.
Wagner, E. et al (1991). Bioconjugate Chem., 2, 226-231.
Wagner, E. et al (1990). Proc. Natl. Acad. Sci. USA, 87,
3410-3414.
Cotten, M. et al (1990). Proc. Natl. Acad. Sci. USA, 87,
4033-4037.
Zenke, M. et al (1990). Proc. Natl. Acad. Sci. USA, 87,
3655-3659.
| Cat
No. |
Size |
| GDS0060 |
32
transfections (24 well plates) |
| GDS0070 |
80
transfections (24 well plates) |
Transfection
Efficiencies of DuoFect™ with various cell types
| Tested
Cell Lines |
Level
of
Luciferase
Expression |
| Human
K562 |
|
| Ewing's
EW-2 Human Sarcoma |
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| HD3
Chicken Erythroblasts |
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| REV-NPB4
Chicken Lymphoblasts |
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| MelJuso
Melanoma |
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| WM
115 Melanoma (Primary Tumor) |
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| WM
266-4 Melanoma (Metastatic) |
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| lgR39
Melanoma (Primary Tumor) |
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| lgR37
Melanoma (Metastatic) |
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| A375(ll)
Melanoma |
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| MelMusa
Melanoma |
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| C32
Melanoma |
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| SK
Mel 28 Melanoma |
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| Colo
320 Colon Carcinoma |
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| LoVo
Colon Carcinoma |
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| SW
403 Colon Carcinoma |
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| SW
948 Colon Carcinoma |
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| LS
174-T Colon Carcinoma |
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| LS
180 Colon Carcinoma |
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| Sk
Col 1 Colon Carcinoma |
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| DLD-1
Colon Carcinoma |
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| Human
HeLa |
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| Human
HepG2 |
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| Human
H9 |
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| Hamster
CHO |
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| Mouse
Ehrlich Ascites |
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| Tested
Cell Lines |
Level
of
Luciferase
Expression |
| Monkey
COS |
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| Rat
H4llEC3 |
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| Rat
lA |
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| Chicken
EGFR-myb Erythroblasts |
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| Chicken
Normal Bone Marrow |
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| H4
Neuroglioma |
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| Human
293 Embryonic Kidney |
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| SSC-9
Epithelial Carcinoma |
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| Kato
Stomach Carcinoma |
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| BT20
Mammary Carcinoma |
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| Human
Primary Lymphocytes |
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| Human
U937 |
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| Human
Burkitt |
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| Human
CCRF CEM |
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Legend
›10.000 RLU g measured with pRSVIuc
1.000 - 10.000 RLU g measured with pRSVIuc
100 - 1.000 RLU g measured with pRSVIuc
10 - 100 RLU g measured with pRSVIuc |
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