amaxa Nucleofector technology flyer - Lonza AG

amaxa xxxxxxxxxxxx Nucleofector research
® technology
Nucleofector ® technology
your link to transfection

Nucleofector ® technology
DNA delivery straight into the
nucleus
Normal human dermal fibroblasts –
neonatal were nucleofected with 2.5 μg
TMR-labeled plasmid DNA encoding
eGFP. After 2 hours, cells were fixed
with 3.5% PFA and analyzed by confocal
microscopy. TMR label is shown
in (A), GFP fluorescence in (B), DAPI
nuclear staining in (C) and a merge of
all three fluorescent labels in (D).
page 2 www.amaxa.com Nucleofector ® technology
The Nucleofector ® technology
is the only reliable non-viral transfection method for a wide
range of difficult-to-transfect cell lines and primary cells.
Established in leading labs
The Nucleofector ® technology has become an established transfection
technology in many labs worldwide. A growing number
of publications illustrates the importance of the technology for
day-to-day research in addressing issues of formerly difficultto-transfect
cell lines and primary cells.
One technology for cell lines and primary cells
Using the Nucleofector technology, cell lines, as well as
primary cells, can be reliably transfected. DNA delivery
straight into the nucleus ensures transfection efficiencies of
up to 90% in both, cell lines and primary cells.
Use of many different substrates
With the Nucleofector technology various nucleic acids
can be easily transfected such as DNA, RNA or siRNA. Transfection
parameters remain the same for DNA, RNA and siRNA
so that no further optimization is required.
Maintenance of functionality
The nucleofection ® procedure has no influence on the cell
functionality. Nucleofected cells show long-term survival in
culture and can easily be differentiated or stimulated.
A
C
B
D

Nucleofector ® technology
new
puC ori
Kanamycin
pmaxGFP
(3486 bp)
SV 40 pA
PCMV
max GFP
Plasmid map of pmaxGFP
Cells transfected with pmaxGFP can be easily analyzed for
transfection efficiency by flow cytometry or fluorescent
microscopy.
Positive controls available
page 3 www.amaxa.com amaxa – your link to transfection
Intron
The two components of the
Nucleofector ® technology
The Nucleofector ® II Device
The Nucleofector II Device delivers unique electrical parameters
that are different from other commercially available electroporation
instruments. Electrical settings are pre-programmed for
each optimized cell type. The integrated chipcard reader allows
quick program and software updates for future applications.
> easy handling motor-driven carousel for automatic cuvette handling
> high convenience large graphic display for easy day-to-day use
> great flexibility software enables the filing of individual programs
> future proof suitable to use together with the 96-well Shuttle
The Nucleofector ® Kits
The Nucleofector Kits contain amaxa specified cuvettes, pipettes,
pmaxGFP, Nucleofector Solution and Supplement. Each
solution and supplement is individually developed for every
primary cell type. For cell lines, five different Nucleofector
Solutions, C, L, R, T and V, are available. All solutions provide a
cell-friendly environment that ensure the highest transfection
results and cell viability. Nucleofector Kits are only functional
in combination with the Nucleofector Device.
Introducing maxGFP
All Nucleofector Kits contain a plasmid encoding maxGFP, a
green fluorescent protein from the copepod Pontellina p.
The new siRNA Test Kit offers the opportunity to easily establish
siRNA in cell lines or adherent primary cells. The kit contains
pmaxGFP and an siRNA duplex directed against maxGFP together
with a detailed protocol. Gene silencing can thus be easily
monitored by decrease of maxGFP expression.
siRNA Test Kit Cat. No. VSC-1001

cell lines
The ideal tool for
transfection of cell lines
The Nucleofector technology is the ideal tool for transfection of
virtually any type of cell line, such as, commonly-used cell lines
or difficult-to-transfect cell lines. High transfection efficiencies
are combined with high cell viability and reproducibility of transfection
results.
> high efficiencies up to 90% gene transfer efficiency with high cell viability
> ideal for siRNA up to 99% transfection efficieny with siRNA duplexes even
in suspension cells
> safe and easy procedure easy and convenient handling
> fast experiments expression within hours - from transfection to analysis in
Visit the amaxa
cell line database
Enter your cell line
of interest or view
complete list
Screen results
one day
page 4 www.amaxa.com Nucleofector ® technology
How to find your cell line of interest
Cell type listed
Click on cell name
Your cell line of interest
Cell type not listed
Use Cell Line Optimization
Nucleofector Kit or contact
amaxa’s Scientific Support
Use recommended
for further advice.
Nucleofector Kit in combination
with the respective Optimized
Protocol or use the settings
optimized by other users.

cell lines
d
Optimization of virtually any cell line in just one experiment
Difficult-to-transfect The advanced Cell Line Optimization Nucleofector Kit is
cell lines an ideal tool for the transfection of your difficult-to-transfect
cell line. It enables you to conveniently determine the optimal
nucleofection parameters of your cell line of interest within one
experiment.
1 Solution L V
program 1 A-020 A-020
program 2 T- 020 T- 020
program 3 T-030 T-030
program 4 X-001 X-001
program 5 X-005 X-005
program 6 L- 0 2 9 L- 0 2 9
program 7 D-023 D-023
Step 1 The cell line of interest is transfected
with the Nucleofector Solutions
L and V in combination with seven
different Nucleofector programs.
Examples of amaxa optimized cell lines
293 ATCC ® CRL-1573 84%
293 (DSMZ) 57%
32D ATCC ® CRL-11346 79%
3T3-L1 (pre-ad) ATCC ® CL-173 73%
A-10 ATCC ® CRL-1476 64%
A20 ATCC ® TIB-208 74%
A-431 ATCC ® CRL-1555 45%
A549 ATCC ® CCL-185 72%
AGS ATCC ® CRL-1739 73%
BA/F3 (DSMZ) 88%
BHK-21 ATCC ® CCL-10 85%
BJ ATCC ® CRL-2522 52%
C2C12 ATCC ® CRL-1772 82%
C6 ATCC ® CCL-107 95%
Caco-2 ATCC ® HTB-37 59%
CCRF-CEM ATCC ® CCL-119 68%
CHO-K1 (DSMZ) 84%
CHO-K1 ATCC ® CCL-61 94%
CHO-suspension (ECACC) 92%
COS-1 ATCC ® CRL-1650 49%
COS-7 (DSMZ) 85%
DU 145 ATCC ® HTB-81 47%
EL4 ATCC ® TIB-39 65%
FDC-P1 ATCC ® CRL-12103 82%
HaCaT (DKFZ) 43%
HCT 116 ATCC ® CCL-247 78%
For an updated cell list and to view customer derived data for approximately 300 additional cell types please go to
www.amaxa.com/celldatabase.
Step 2 The Nucleofector Solution and
program which result in highest transfection
efficiencies with lowest mortality
are selected.
HeLa ATCC ® CCL-2 70%
HeLa (DSMZ) 57%
HepG2 ATCC ® HB-8065 64%
HL-60 ATCC ® CCL-240 50%
HT-29 ATCC ® HTB-38 51%
HT-1080 ATCC ® CCL-121 74%
HuT 78 ATCC ® TIB-161 53%
IMR-90 ATCC ® CCL-186 51%
JURKAT ATCC ® TIB-152 54%
JURKAT (DSMZ) 63%
K562 ATCC ® CCL-243 89%
K562 (DSMZ) 76%
KG-1 ATCC ® CCL-246 70%
LNCaP ATCC ® CRL-1740 70%
MCF-7 ATCC ® HTB-22 71%
MDA-MB-231 ATCC ® HTB-26 80%
MDA-MB-453 ATCC ® HTB-131 54%
MDA-MB-468 ATCC ® HTB-132 60%
MDCK ATCC ® CCL-34 73%
MDCK II (ECACC) 80%
MOLT-4 ATCC ® CRL-1582 55%
NALM-6 (DSMZ) 64%
NB-4 (DSMZ) 71%
NCI-H1299 ATCC ® CRL-5803 99%
Neuro-2a ATCC ® CCL-131 76%
NIH/3T3 ATCC ® CRL-1658 84%
page 5 www.amaxa.com amaxa – your link to transfection
2
> > >
3
V Solution
Step 3 A further fine tuning of the
nucleofection conditions can be performed
with the help of our Scientific
Support Team.
+
> > >
NIH-3T3 (DSMZ) 68%
NRK ATCC ® CRL-6509 44%
NS0 (ECACC) 83%
P19 ATCC ® CRL-1825 85%
PC-12 ATCC ® CRL-1721 34%
PC-3 ATCC ® CRL-1435 81%
PANC-1 ATCC ® CRL-1469 68%
Raji ATCC ® CCL-86 58%
Ramos ATCC ® CRL-1596 27%
RAW 264.7 ATCC ® TIB-71 65%
RBL-1 ATCC ® CRL-1378 84%
S49 ATCC ® TIB-36 81%
Saos-2 ATCC ® HTB-85 82%
SH-SY5Y ATCC ® CRL-2266 82%
SK-OV-3 ATCC ® HTB-77 89%
SK-N-SH ATCC ® HTB-11 85%
SW480 ATCC ® CCL-228 60%
T-47D ATCC ® HTB-133 51%
THP-1 ATCC ® TIB-202 67%
U-2 OS ATCC ® HTB-96 98%
U-937 ATCC ® CRL-1593.2 56%
U-87 MG ATCC ® HTB-14 43%
U266BI ATCC ® TIB-196 86%
Vero ATCC ® CCL-81 79%
WEHI-231 ATCC ® CRL-1702 77%

primary cells
d
Nucleofection of primary cells
In the past, the lack of efficient and easy transfection methods
for primary cells restricted experiments mainly to cell lines.
However, the artifical nature of cell lines limits the significance
of experimental results. Today, the isolation and culture of primary
cells is well-established. With introduction of the unique
Nucleofector technology the last obstacle, the efficient non-viral
transfection of such cell types, was overcome in 2001. The
technology is now a well-established method worldwide.
Researchers are now able to address open questions of their
specific research area in many primary cell types which reflect
a much more relevant physiological system than any cell line
could ever be.
> high efficiencies up to 90% gene transfer efficiency with high cell viability
> no optimization ready-to-use kits for a multitude of cell types
> reproducible results Optimized Protocols for each cell type with details on cell
isolation and culture
> ideal for primary cells direct transport of nucleic acids into the nucleus thus
Functional CD2 silencing in primary
human T cells
Primary human T cells were transfected
with 1.4 μg siRNA targeted to
CD2 using the Human T Cell Nucleofector
Kit. 44 hours post nucleofection,
cells were stained with anti
CD2-FITC antibody (A) or anti CD4-
PE antibody (B) and analyzed by
flow cytometry. Controls included
nucleofection of 1.4 μg control siRNA
and addition of CD2 siRNA without
nucleofection.
page 6 www.amaxa.com Nucleofector ® technology
enabling transfection even of non-dividing cells
siRNA transfection in primary cells -
up to 99% transfection efficiency
The Nucleofector technology ensures efficient delivery of siRNA
even in primary cells enabling strong knock-down effects. High
cell viability of more than 85% is maintained even with higher
amounts of siRNA.
counts
200
160
120
80
40
0
10 0
A
CD2-FITC
10 1
For further information about nucleofection of siRNA in primary cells and cell lines, go to www.amaxa.com/RNAi
or contact our Scientific Support Team.
10 2
10 3
10 4
counts
300
240
180
120
60
0
10 0
CD4-PE
CD2 siRNA control siRNA
no siRNA + nucleofection
B
10 1
10 2
10 3
10 4

primary cells
d
Transfection results you can trust
Examples for primary cell Transfection efficiency*
Nucleofector Kits
Dendritic cells, human 49%
cancer research HUVEC 90%
MEF 43%
etc.
Cardiomyocytes, rat 35%
cardiovascular research HMVEC-L 52%
MSC, human 47%
etc.
Dermal fibroblasts, human 89%
dermatology Keratinocytes, human 53%
etc.
B cells, human 36%
immunology Macrophages, mouse 47%
Macrophages, human 59%
T cells, human 64%
T cell, mouse 39%
etc.
Neurons, mouse 58%
neurobiology Neurons, rat cortical 59%
Neural stem cells, mouse 60%
etc.
Hepatocytes, mouse 54%
gastroenterology Hepatocytes, rat 52%
* Data show examples of average transfection efficiencies given as % positive viable cells. Cells were either transfected with a plasmid encoding maxGFP, eGFP
or H-2K k , a mouse MHC class I molecule. Analysis was performed by either flow cytometry or fluorescence microscopy. Cell viability varies between 50-90%.
Protect your cells from mycoplasma
amaxa offers new antibiotic formulations specifically developed to protect cell lines or sensitive
primary cells against mycoplasma, gram+ and gram- bacteria as well as fungi.
Primocin for primary cells Cat. No. VZA-1021
Normocin for cell lines Cat. No. VZA-1001
Plasmocin for mycoplasma cure Cat. No. VZA-1012
For more information about the new cell culture antibiotics, see www.amaxa.com/antibiotics.
page 7 www.amaxa.com amaxa – your link to transfection

WKB-1001_01.2006
d
For an up-to-date
overview or more
detailed information
about amaxa products,
please see
www.amaxa.com/
productlist.
Cat.No.
AAD-1001 Nucleofector ® device
VPG-1006 Astrocytes, mouse
VPG-1007 Astrocytes, rat
VPA-1001 B cells, human
Ordering information
Nucleofector ® Kits for primary cell types
VPE-1002 Cardiomyocytes, rat neonatal
VPA-1003 CD34+ cells, human
VPF-1001 Chondrocytes, human
VPA-1004 Dendritic cells, human
VPD-1001 Dermal fibroblasts, human
VPD-1006 Embryonic fibroblasts, mouse, Starter Kit
VPD-1004 Embryonic fibroblasts, mouse, Kit I
VPD-1005 Embryonic fibroblasts, mouse, Kit II
VPH-1001 Embryonic stem cells, mouse
VPB-1001 Endothelial cells, human coronary artery (HCAEC)
VPB-1003 Endothelial cells, human microvascular (HMVEC-L)
VPB-1002 Endothelial cells, human umbilical vein (HUVEC)
VPK-1001 Epithelial cells, human coronary artery
VPK-1002 Epithelial cells, human mammary
VPK-1003 Epithelial cells, human prostate
VPL-1002 Hepatocytes, mouse
VPL-1003 Hepatocytes, rat
VPD-1002 Keratinocytes, human
VPA-1008 Macrophages, rat
VPA-1009 Macrophages, mouse
VPA-1007 Monocytes, human
VPD-1003 Melanocytes, human epidermal neonatal (NHEM)
VPE-1001 Mesenchymal stem cells, human
VPA-1005 Natural killer cells, human (NK)
VPG-1004 Neural stem cells, mouse
VPG-1005 Neural stem cells, rat
VPG-1002 Neurons, chicken
VPG-1001 Neurons, mouse
VPG-1003 Neurons, rat
VPG-1009 Oligodendrocytes, rat
VPC-1001 Smooth Muscle Cells, human aortic (AoSMC)
VPA-1002 T cells, human
VPA-1006 T cells, mouse
Basic Nucleofector ® Kits
VPI-1001 Mammalian endothelial cells
VPI-1002 Mammalian fibroblasts
VPI-1003 Mammalian neural cells
VPI-1004 Mammalain smooth muscle cells
VPI-1005 Mammalian epithelial cells
Nucleofector ® Kits for cell lines
VCO-1001 Cell Line Optimization Nucleofector ® Kit
VCA-1004 Cell Line Kit C
VCA-1005 Cell Line Kit L
VCA-1001 Cell Line Kit R
VCA-1002 Cell Line Kit T
VCA-1003 Cell Line Kit V
amaxa's Nucleofector ® Process, Nucleofector ® Device and Nucleofector ® Solutions are covered by PCT Applications PCT/EP01/07348,
PCT/DE02/01489, PCT/DE02/01483, and other pending patents, and domestic or foreign applications corresponding thereto.
amaxa, Nucleofector, nucleofection, maxGFP and 96-well Shuttle are trademarks of amaxa GmbH.
The CMV promoter is covered under U.S. Patents 5,168,062 and 5,385,839 and its use is permitted for research purposes only. Any other use of
the CMV promoter requires a license from the University of Iowa Research Foundation, 214 Technology Innovation Center, Iowa City, IA 52242.
ATCC ® and the ATCC Catalog Marks are trademarks of ATCC used under License.
amaxa GmbH, Europe/World amaxa Inc., USA
Scientific Support Scientific Support
+49 (0)221-99199-400 (240) 632-9110
scientific-support@amaxa.com www.amaxa.com scientific-support.US@amaxa.com