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OZ Biosciences – PolyMag Neo – v2 Page 1 of 10
PolyMag Neo Results
PolyMag Neo is based on OZ Biosciences Magnetofection Technology. It was specifically developed to achieve high transfection efficiency (percentage of cells transfected) combined with superior transgene expression level. PolyMag Neo represents the latest development of our magnetic nanoparticles based transfection reagents. In this way, PolyMag Neo is the ideal transfection reagent in a wide variety of cells.
Main features are:
• High transgene expression • Good transfection efficiency • Multipurpose (various types of nucleic acids) • Universal (primary cells and cell lines) • Simple, Ready-to-use and Rapid • Non toxic • Compatible with and without serum-containing culture media
Nucleic acid types
PolyMag Neo Transfection Reagent is suitable for all type of nucleic acids including: plasmid DNA, siRNA, oligonucleotides, linearized DNA, double stranded RNA, mRNA, shRNA.
Cell Types
PolyMag Neo is suitable for numerous cells. This reagent has been successfully tested on several cells (see Table 1). If a particular cell type is not listed, this does not imply that PolyMag Neo is not going to work. OZ Biosciences is maintaining an updated list of cells successfully tested that is available on the website: www.ozbiosciences.com. You can also submit your data to [email protected] so we can update this list and give you all the support you need.
PolyMag Neo transfection efficiency on various cell lines
Several cell lines (1x105 cells/ well) were transfected with 0.5 µg of pEGFP plasmid DNA per well in a 24-well plate. Transfections were performed with 0.5 µL per well of PolyMag Neo transfection reagent. Percentage of transfected cells were measured 24h post transfection by flow cytometry. Table 1: Example of cells successfully transfected with PolyMag Neo Transfection Reagent.
Cells Cell Type Species % Transfected
Cells
Transgene Expression Level
293, 293T Kidney Human 90 % +++ A549 Non-small cell lung carcinoma Human 45-55 % ++ B65 Neuroblastoma Rat 30-40 % + BEAS-2B Bronchial Epithelial Human 55-65 % +++ BHK-21 Kidney Hamster 65-75 % +++ C6 Glial Tumor Rat 25-35 % + CHO, CHO-K1 Ovary (epithelial like) Chinese Hamster 85 % +++ COS-1 , COS-7 Kidney Green Monkey 80 % +++
OZ Biosciences – PolyMag Neo – v2 Page 2 of 10
HaCat Keratinocyte Human 40-50 % + HEK293 Kidney Human 90 % +++ HeLa, HeLa-S3 Cervix carcinoma Human 70-80 % +++ Jurkat * Acute T cell lymphocyte Human < 5 % ND L929 Fibrosarcoma cells Mouse 30 % + MDCK Kidney Dog 45-55 % +++ MS-5 Bone marrow stroma cells mouse 80-90% +++ NIH-3T3 Fibroblasts Mouse 60 % ++ RAW Macrophage Mouse 5-15 % + SH-SY5Y Neuroblastoma Human < 5 % + U87 Glioma Human 70 % + Vero Kidney Green Monkey 25-35 % +++
PRIMARY CELLS
HUVEC Human 40-50 % Chondrocytes Porcine ND Fibroblasts Mouse ND
* suspension cells ND: Not determined +++: ≥ 200 ng transgene expression level in a well of 24-well plate ++: 100 to 200 ng transgene expression level in a well of 24-well plate +: ≤ 100 ng transgene expression level in a well of 24-well plate
Efficiency of PolyMag Neo on several cell lines
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BEAS MDCK BHK21 HeLa A549 HEK293 MS-5 HaCat NIH3T3 C6 B65 U87 VERO
% o
f T
rans
fect
ed C
ells
Cell Line
PolyMag Neo Transfection Efficiency
Cells (1x105) were transfected with 0.5 µg / well of pEGFP plasmid DNA in 24-well plates. Transfections were performed with 0.5 µl / well of polyMag Neo reagent. Percentage of transfected cells were measured 24h post transfection by flow cytometry. For an updated list of cells successfully transfected, please visit our website: www.ozbiosciences.com.
OZ Biosciences – PolyMag Neo – v2 Page 3 of 10
GFP in different cells transfected with PolyMag Neo Cells (1x105) were transfected with 0.5 µg / well of pEGFP plasmid and 0.5 µL of PolyMag Neo reagent in 24-well plates. EGFP expression was monitored 24 h after transfection by fluorescence microscopy.
BHK21
HeLa
U87
VERO-10A1 MDCK
BEAS-2B
HEK293 MS-5
OZ Biosciences – PolyMag Neo – v2 Page 4 of 10
β-Galactosidase expression in different cell lines transfected with PolyMag Neo Cells (1x105) were transfected with 0.5 µg / well of pLacZ plasmid and 0.5 µL of PolyMag Neo reagent in 24-well plates. β-Galactosidase expression was revealed 24 h after transfection using OZ Biosciences’ X-Gal staining kit (catalog number GX-10003).
PolyMag Neo efficiency compared to other transfection reagents
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MDCK B65
% o
f Tr
ansf
ecte
d Ce
lls
cell line
Comparison with other transfection reagents
PolyMag Neo
Reagent FH
Reagent DG
Reagent IB
Reagent L2
HEK293 MS-5
OZ Biosciences – PolyMag Neo – v2 Page 5 of 10
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Hela BEAS Cos-7 NiH3T3
Re
lati
ve A
mou
nt
of
GFP
(%)
Comparison with other transfection reagents - GFP
PolyMag Neo
Reagent FH
Reagent DG
Reageant LX
cell line
Cells (1x105) were transfected with 0.5 µg / well of pEGFP plasmid and 0.5 µL of PolyMag Neo reagent in 24-well plates. Other transfection reagents were used as recommended by manufacturers. GFP expression and % of transfected cells were revealed 24 h after transfection using flow cytometry. Results are expressed in percentage of relative values.
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HeLa BEAS
Rel
ati
ve A
mou
nt o
f β-
gala
ctos
ida
se (%
)
Comparison with other transfection reagents - β-galactosidase
PolyMag Neo
Reagent FH
Reagent DG
Reagent IB
Reagent LX
cell line
Cells (1x105) were transfected with 0.5 µg / well of pLac-Z plasmid and 0.5 µL of PolyMag Neo reagent in 24-well plates. Other transfection reagents were used as recommended by manufacturers. β-Galactosidase was revealed 24 h after transfection using OZ Biosciences’ ONPG β-galactosidase assay kit (catalog number G010001). Results are expressed in percentage of relative values.
Cell line
OZ Biosciences – PolyMag Neo – v2 Page 6 of 10
Relative amount of protein expression between PolyMag Neo and PolyMag
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VERO BEAS A549 Cos7 HaCat NIH-3T3 U87 MS-5
Re
lati
ve A
mo
un
t og
GFP
(A
U)
superior gene expression level-GFP
PolyMag Neo
PolyMag
cell line
Cells (1x105) were transfected with 0.5 µg or 1 µg / well of pEGFP plasmid DNA in 24-well plates for PolyMag Neo and PolyMag respectively. 0.5 µL or 1 µL per well of PolyMag Neo or PolyMag were used. GFP average intensity of each well was measured 24 h after transfection by flow cytometry and results are expressed as relative values.
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600
MDCK BHK21 HeLa C6 CHO B65 HEK293
Re
lati
ve A
mo
un
t of β
-ga
lact
osi
da
se (
ng
)
superior gene expression level-β galactosidase
PolyMag Neo
PolyMag
cell line Cells (1x105) were transfected with 0.5 µg or 1 µg / well of pLacZ plasmid DNA in 24-well plates for PolyMag Neo and PolyMag respectively. 0.5 µL or 1 µL per well of PolyMag Neo or PolyMag were used. β-Galactosidase expression was revealed 24 h after transfection using OZ Biosciences’ ONPG β-galactosidase assay kit (catalogue number G010001). β-galactosidase amount of each well was measured by spectrometry and results are expressed in ng of .
OZ Biosciences – PolyMag Neo – v2 Page 7 of 10
Cell Viability
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Non Transfected PolyMag PolyMag Neo Reagent DG Reagent IB Reagent G2
Per
cent
age
of L
ivin
g C
ells
(%
)
Effect of various transfection reagents on cell viability
HeLa cells (1x105) were transfected with 0.5 µg of pLacZ plasmid and 0.5 µL of PolyMag Neo per well in a 96-well plate. Other transfection reagents were used as recommended by manufacturers.
Optimization of DNA Amount
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0.25 µg 0.5 µg 1 µg
% o
f Tra
nsfe
cted
Cel
ls
DNA Quantity
MDCK
HeLa
C6
OZ Biosciences – PolyMag Neo – v2 Page 8 of 10
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0.25 µg 0.5 µg 1 µg
% o
f Tra
nsfe
cted
Cel
ls
DNA Quantity
BEAS-2B
A549
HEK293
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1400
0.25 µg 0.5 µg 1 µg
GFP
am
ount
(AU
) BEAS-2B
C6
A549
CHO
DNA quantity
Cells (1x105) were transfected with 0.25 µg, 0.5 µg and 1 µg / well of pEGFP plasmid DNA in 24-well plates. PolyMag Neo transfections were performed with 0.25 µl / well, 0.5 µl / well and 1 µl / well of reagent. GFP expression level and % of transfected cells were monitored by flow cytometry.
OZ Biosciences – PolyMag Neo – v2 Page 9 of 10
Optimization of PolyMag Neo/DNA Amount
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0.2 µg 0.4 µg 0.8 µg 1.6 µg
Am
oun
t of β
-gal
acto
sida
se (
ng)
DNA quantity
A549 cells
A549 cells (15x103) were transfected with 0.2 µg, 0.4 µg, 0.8 µg and 1.6 µg / well of pLacZ plasmid and 0.2 µL, 0.4 µL, 0.8 µL and 1.6 µL of PolyMag Neo reagent in 96-well plates. β-Galactosidase expression was revealed 24 h after transfection using OZ Biosciences’ ONPG β-galactosidase assay kit (catalog number G010001). β-galactosidase amount of each well was measured by spectrometry.
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0.4 µL 0.8 µL 1.6 µL 3.2 µL
Am
ount
of β
-ga
lact
osid
ase
(ng)
PolyMag Neo quantity
A549 cells
A549 cells (15x103) were transfected with 0.8 µg / well of pLacZ plasmid and 0.4 µL, 0.8 µL, 1.6 µL and 3.2 µL of PolyMag Neo reagent in 96-well plates. β-Galactosidase expression was revealed 24 h after transfection using OZ Biosciences’ ONPG β-galactosidase assay kit (catalog number G010001). β-galactosidase amount for each well was measured by spectrometry.
OZ Biosciences – PolyMag Neo – v2 Page 10 of 10
Gene Silencing with PolyMag Neo
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90,0
1nM 5nM 20nM
GFP
inhi
biti
on (
%) Hela GFP
Cos-7 GFP
Final siRNA concentration (nM)
PolyMag Neo/siRNA-GFP
HeLa and Cos-7 (5x104 cells/well) cells stably transfected with a GFP plasmid DNA were treated with 1 nM, 5 nM, and 20 nM of siRNA GFP per well in a 24-well plate. Transfections were performed with 1 to 3µL per well of PolyMag Neo transfection reagent. GFP expression was analysed 48 h post-transfection by flow cytometry.
Hela GFP control PolyMag Neo/Si-GFP 1nm PolyMag Neo/Si-GFP 20nm