| |
Transfection
Reagents |
| TROJENE™
- just add to your cells without
changing the medium. |
Avanti introduces the newest, most effective
alternative to
viral vector technology
|
Developed by IC-Vec
Ltd in association with the Imperial
College Genetic Therapies Centre.
| Product |
Catalog Number |
|
| Trojene |
890308P |
Pricing
|
Protocol
Compare Trojene with the existing market leaders.
Transfection
Reagent
Product "T"1 |
 |
 |
Trojene
Transfection
Procedure For
"Maximum Level
of Transfection"4 |
Transfection
Reagent
Product "L"2 |
 |
 |
Trojene
Transfection
Procedure For
" Minimal
Handling"5 |
Transfection
Reagent
Product "F"3 |
 |
 |
Trojene
Transfection
Procedure For
" Minimal
Toxicity"6 |
The Experiment followed this protocol: 96-well plate with HeLa cells seeded
at a density of 10,000 the day before the experiment. 0.5 µg DNA per
well in all cases. All transfections were carried out overnight. In the
morning, the media were removed and the cells supplemented with 250 µl
of fresh complete medium. The plasmid DNA encodes for the EGFP, the expression
of the gene is under the control of the CMV promoter. The photos were taken
24 h post-transfection.
Experiment performed by Iratxe Puebla, Imperial College London
1 5 µl reagent (1mg/ml) used,
complex prepared according to manufacturer's instructions, and brought
to 125 µl using complete DMEM. Then added to cells containing 125
µl complete DMEM (growth medium).
2,3 1.25 µg reagent used, complex prepared
in DMEM to a total volume of 25 µl. Final volume per well was 250
µl, medium
DMEM.
4 Trojene operating protocol 1.
5 Trojene operating protocol 2.
6Trojene operating protocol 3.
The experiment was carried out in a 48 well plate with 0.5 µg of DNA
per well in all cases (cell line and incubation time specified on the graph).
The plasmid encodes for beta-galactosidase under CMV promotor. The cells
were assayed for gene expression 24h post-transfection using a standard
chemio-luminescent assay.
We invite you to test this exciting,
new reagent in your laboratory.
The experiment was carried out on 3T3 cells in a 48 well plate with 0.25
µg of DNA per well in all cases. The plasmid encodes for luciferase
under CMV promotor. The cells were assayed for gene expression 24h post-transfection
using a standard luminescent assay.
Trojene and Product "E" were transfected in Medium containing
10% serum.
Product "L" was transfected in Optimem.
References
- Patent number: EP 0918790; N09745442 (polycationic sterol derivatives
as transfection agents).
Publications referring to CD AN : (T rojene)
- Keller, M., M. R. Jorgensen, E. Perouzel, A. D. Miller. (2003). Thermodynamic
Aspects and Biological Profile of CDAN/DOPE and DC-Chol/DOPE Lipoplexes.
Biochemistry, In Press
- Tagawa, T., M. Manvell, N. Brown, M. Keller, E. Perouzel, K. D. Murray,
R. P. Harbottle, M. Tecle, F. Booy, M. C. Brahimi-Horn, C. Coutelle,
N. Lemoine, R., E. W. F. W. Alton and A.
D. Miller. (2002). Gene Therapy, 9, 564.
- Cooper, R.G., C. J. Etheridge, L. Stewart, J. Marshall, S. Rudginsky,
S. H. Cheng, A. D. Miller. (1998). Chemistry-a European Journal . 4,
137.
- Stewart, L., M. Manvell, E. Hillery, C. J. Etheridge, R. G. Cooper,
H. Stark, M. van-Heel, M. Preuss, E. Alton and A. D. Miller. (2001).
- Journal of the Chemical Society-Perkin Transactions 2, 624.
A. J. Geall, A.J., M. A. W. Eaton, T. Baker, C. Catterall, I. S. Blagbrough.
(1999).
Febs Letters, 459, 337.
- A. J. Geall, A.J., R. J. Taylor, M. E. Earll, M. A. W. Eaton, I. S.
Blagbrough. (2000).
Bioconjugate Chemistry, 11, 314; and references therein.
- Keller M, Harbottle RP, Perouzel E, Colin M, Shah I, Rahim A, Vaysse
L, Bergau A, Moritz S, Brahimi-Horn C, Coutelle C, Miller AD. (2003).
Nuclear Localisation Sequence Templated Nonviral Gene Delivery Vectors:
Investigation of Intracellular Trafficking Events of LMD and LD Vector
Systems.
Chembiochem. 2003 Apr 4;4(4):286-98. [PubMed]
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