Brochure "TurboFect™ in vitro Transfection Reagent ... - Biocenter

TurboFect
in vitro
Transfection Reagent
P26

Ready-to-use reagent
TurboFect in vitro Transfection Reagent is a highly efficient, easy-to-use, non-immunogenic transfection reagent.
It is a sterile solution of a proprietary cationic polymer in water. The polymer forms compact, stable, positively charged
complexes with DNA. These complexes protect DNA from degradation and facilitate gene delivery into eukaryotic cells.
Features:
• High transfection efficiency of a wide
variety of cell types
• No optimization required for most cell lines
• Excellent transfection efficiency in the presence
or absence of serum
• Minimal cytotoxicity
• Ready-to-use - no need to reconstitute,
dilute or manipulate
TurboFect – mechanism of action
Applications:
• Stable and transient DNA transfection
• Co-transfection
• Transfection of RNA transcripts
• Transfection of:
– primary and secondary cell lines
– adherent and suspension cells
– differentiated and undifferentiated cells
Cell membrane
Cytoplasm
H 2 O
Endosome rupture
H 2 O
DNA
Lysosome
TurboFect / DNA
complexes
Endosome
H + Cl – Nucleus
Endosome/lysosome
DNA
Endosome
Lysosome
• TurboFect interacts with DNA to form small, stable, highly diffusible
complexes which are readily endocytosed.
• “Proton-sponge” effect of TurboFect buffers endosomal pH by
provoking massive proton accumulation and passive chloride influx.
• Rapid osmotic swelling causes endosomal rupture, allowing translocation
of DNA to the nucleus.
Intracellular localization of FITC-labeled DNA-TurboFect
complexes 4 hours after transfection with TurboFect .
Cell nuclei were stained with DAPI and lysosomes were labeled
with PE-conjugated anti-LAMP1 antibody. Images were taken
using a laser confocal microscope and represent 4 combined
cross-sections through the nucleus.
TurboFect – fast and simple transfection procedure
Transfection with
TurboFect Cationic Lipids Calcium Phosphate
Prepare transfection solutions
Mix TurboFect with DNA Mix transfection reagent with DNA Mix transfection reagent with DNA
Prepare cells by changing to
serum-free media
Prepare cells by changing to
serum-free media
Add TurboFect -DNA complex and
incubate
Add transfection reagent-DNA
complex and incubate
Add transfection reagent-DNA
complex and incubate
Wash off transfection reagent to
prevent cytotoxic effects
Wash off transfection reagent to
prevent cytotoxic effects
Replace with fresh serum-enriched
media
Replace with fresh serum-enriched
media
Assay for gene expression Incubate and then assay for gene
expression
Incubate and then assay for gene
expression
3 steps 6 steps 7 steps
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High transfection efficiency
High transfection efficiency of a wide variety of cell types
100
GFP positive cells (%)
80
60
40
20
0
B50
calu1
CHO
COLO
Cos-7
HEK293
HeLa
HeLa S3
Hep2C
HepG2
L929
RAW264
MDCK
NIH3T3
Sp2/Ag14
WEHI
Transfection of different cell lines with TurboFect in vitro Transfection Reagent.
Cells were transfected with plasmid DNA encoding eGFP. Transfection efficiency was analyzed by flow cytometry.
TurboFect – superior transfection reagent
GFP positive cells (%)
100
80
60
40
20
0
HEK293
TurboFect ExGen 500 Vendor A Vendor B Vendor C Vendor D Vendor E Vendor F NC
4000
3000
2000
1000
0
fluorescence intensity
GFP positive cells (%)
60
50
40
30
20
10
0
TurboFect ExGen 500 Vendor A Vendor B Vendor C Vendor D Vendor E Vendor F NC
HepG2
1500
1250
1000
750
500
250
0
fluorescence intensity
Comparison of TurboFect in vitro Transfection Reagent with tranfection reagents from other vendors.
Cells were transfected with plasmid DNA encoding eGFP. Transfections were performed according to manufacturer’s recommendations and
analyzed by flow cytometry.
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The only transfection reagent you need
TurboFect is effective even in
hard-to-transfect cell lines
Reliable co-transfection
EGFP expression in primary mouse bone marrow derived
dendritic cells transfected with TurboFect in vitro
Transfection Reagent.
Triple transfection of HeLa cells.
HeLa cells were co-transfected with a 1:1:1 mixture of plasmids
encoding eGFP localized in the cytoplasm, eCFP with a nuclear
localization signal and monomeric DsRed protein with membrane
localization signal. The cells were transfected with TurboFect in vitro
Transfection Reagent.
Cells successfully transfected with TurboFect in vitro Transfection Reagent include:
Permanently growing cell lines
• B50 rat nervous tissue neuronal cells
• BHK21 baby hamster kidney cells
• Calu1 human lung epidermoid carcinoma cells
• CHO chinese hamster ovary cells
• COLO human colon adenocarcinoma cells
• Cos-7 african green monkey kidney cells
• HEK293 human embryonic kidney cells
• HeLa human cervix adenocarcinoma cells
• HeLa S3 human cervix carcinoma cells
• Hep2C human larynx carcinoma cells
• HepG2 human hepatoma cells
• Jurkat human leucaemic T cells
• L929 mouse connective tissue fibroblasts
• RAW264 mouse leucaemic monocyte-macrophage cells
• MDCK Madin Darby Canine Kidney cells
• NIH3T3 mouse embryo fibroblasts
• Sp2/Ag14 mouse myeloma cells
• WEHI mouse B cell lymphoma cells
Primary cell cultures
• Rat fibroblasts
• Mouse bone marrow derived dendritic cells
• Mouse bone marrow derived macrophages
• HLF human lung fibroblasts
For cell line updates, see
www.fermentas.com/catalog/transfection
Product Cat. # Size
TurboFect in vitro Transfection Reagent #R0531 1 ml (delivers 500 µg of DNA)
PureExtreme ® is registered Fermentas trademark.
TurboFect is Fermentas trademark.
ExGen is a trademark of Euromedex.
Fermentas – a member of your research team www.fermentas.com
© Fermentas 2009