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LyoVec New formulation
LyoVec is the first lyophilized cationic lipid-based transfection reagent. The major constituent of LyoVec is the phosphonolipid DTCPTA, which
is coupled with DiPPE, a neutral lipid that helps destabilizing membrane bilayers, therefore increasing the in vitro transfection efficiency of LyoVec .
Features and Benefits
Rapid and Simple-to-use
- Hands-on time: No more than 15 min
- No preparation required the day prior transfection
Optimized Procedure
- Works similarly at various lipid/DNA ratios
- No optimization of transfection conditions
- Soluble in water up to 5X
Maximum Transfection Efficiency
- Transfects with high efficiencies a broad spectrum of mammalian cells,
including primary cells and non-adherent cells
- Effective for transient and stable transfections
Minimal Cytotoxicity
- Works in the presence of serum
- No need to wash cells after transfection
Increased Stability
- Stable over one year when lyophilized
- Stable up to six months when rehydrated
Long shelf-life of pDNA/LyoVec Complexes
In contrast with other cationic lipids, plasmid DNA (pDNA)/
LyoVec complexes remain fully active for transfection for at least two
months at 4°C. Thus, preparation of large volumes of complexes can be
made and reused repeatedly, saving you time.
Structure
DTCPTA
Di-tetradecylphosphoryl-N,N,N-trimethylmethanaminium chloride
1. Floch et al. 1997. Cationic phosphonolipids as non viral vectors for DNA transfection
in hematopoietic cell lines and CD34+ cells. Blood Cells, Molec.& Diseases 23: 69-87.
2. Guillaume-Gable et al. 1998. Cationic phosphonolipids as nonviral gene transfer
agents in the lung of mice. Hum. Gene Ther. 9: 2309-2319
Contents and Storage
LyoVec is provided as a lyophilized powder, sterile and packaged in sealed
2 ml glass vials (4 or 9 vials). After reconstitution, each vial yields 2 ml of
transfection reagent allowing 40 transfections of 1-4 x 10 5 cells/well in a 12well
plate. Each order of LyoVec comes with 1 vial of control LyoVec /GFP-
LacZ complex. LyoVec is shipped at room temperature. Upon receipt, store
at 4°C. LyoVec is stable up to 1 year when properly stored.
PRODUCT QUANTITY CAT. CODE
LyoVec 8 ml (160 reactions)
18 ml (360 reactions)
lyec-12
lyec-22
Percentage of b-Gal-expressing cells
Invivo
LyoV
LyoVec Procedure
STEP 1 - LyoVec Reconstitution
Reconstitute LyoVec with 2 ml sterile
H2O or PBS
Note: Reconstituted LyoVec can be stored
at 4°C for 6 months.
STEP 2 - Transfection of Adherent and
Suspension Cells
1-10 μl pDNA (1-3 μg) + 100 μl LyoVec
-> 20’ at room temperature
Note: Bulk quantities of pDNA/LyoVec
complexes can be prepared for multiple
wells.
1 ml cell suspension (0.25-1.10 6 cells) +
100 μl pDNA/LyoVec complexes ->
mix gently. Incubate at 37°C in 5% CO2
for 24-72 hours before testing transgene
expression or applying antibiotic
selection.
DiPPE
1,2-Diphytanoyl-sn-Glycero-3-Phosphoethanolamine
Transfection efficiencies of LyoVec : 2 to 8 x 10 5 cells per 6-well plates were
transfected with 1 μg pVITRO2-GFP/LacZ complexed with 100 μl LyoVec in 2 ml
of culture medium containing 10% FBS. LacZ expression was assessed 48h posttransfection.
Note: Reagent X is one of the most utilized transfection reagents on
the market. Transfection was performed according to the manufacturer’s protocol.
www.invivogen.com/transfection-reagents 19
CELL CULTURE & TRANSFECTION 1