TransIT

Transcription

TransIT

2013-14 Transfection Products
www.TheTransfectionExperts.com
ONE FOCUS: TRANSFECTION
Transfection is a type of macromolecular delivery in which large, negatively charged nucleic acids are
delivered through cellular membranes and into the intracellular compartments where they can exert
their function. Almost two decades after its founding, transfection continues to be the focal point and
passion of Mirus Bio.
Every transfection product offered by Mirus Bio is the result of scientific discovery and development from an in-house team of organic chemists, molecular biologists and cell biologists. When
you contact Mirus Bio, you are speaking with a scientist with true expertise in the development,
manufacture, and performance of the product.
MOST RECENT TRANSFECTION BREAKTHROUGHS
2013: TransIT-X2™ Dynamic Delivery System—For superior delivery of plasmid DNA and siRNA
TransIT®-BrCa Transfection Reagent—The first breast cancer cell transfection reagent
2011: TransIT®-3D Transfection Reagent—For use in 3D cell culture scaffolds
2010: TransIT-PRO® Transfection Kit—Large-scale, high yield protein production
2009: TransIT®-2020 Transfection Reagent—High performance broad spectrum transfection
2008: Ingenio® Electroporation Kits & Solution—Versatile, multi-platform electroporation solution
OUR PROMISE TO YOU: SERVICE THROUGH EXPERTISE
Order Online 24/7 (Credit Cards or Purchase Orders)
www.mirusbio.com
Technical and Customer Service
Hours of Operation: 8:00 AM–4:00 PM Central Time, Monday–Friday
Technical Support Email: [email protected]
Customer Service Email: [email protected]
Phone: 888.530.0801 (toll free in the U.S.) or +1.608.441.2852
Fax: +1.608.441.2849
U.S. Distribution
Direct: www.mirusbio.com
Fisher Scientific: www.fishersci.com
International Distribution
For a complete list of our worldwide
distributors, please visit: www.mirusbio.com
and contact the distributor in your region.
Mail
Mirus Bio LLC
Attention: Customer Service
545 Science Drive
Madison, WI 53711 USA
Request FREE Samples
Start with our Reagent Agent® transfection database to
determine the best solution for your transfection needs:
www.mirusbio.com/ReagentAgent
Two ways to request a sample:
1. Visit us at www.TheTransfectionExperts.com
2. Call us at 888.530.0801
CONTENTS
DNA/RNA TRANSFECTION
BROAD SPECTRUM DNA & siRNA/miRNA
NEW! TransIT-X2™ Dynamic Delivery System....................................................................2-5
DNA TRANSFECTION
BROAD SPECTRUM DNA
TransIT®-2020 Transfection Reagent..................................................................................6-7
TransIT®-LT1 Transfection Reagent.....................................................................................8-9
CELL LINE SPECIFIC
TransIT®-293 Transfection Reagent..................................................................................... 10
TransIT®-3T3 Transfection Kit.............................................................................................. 10
NEW! TransIT®-BrCa Transfection Reagent .................................................................10 & 12
TransIT®-CHO Transfection Kit............................................................................................. 10
TransIT®-COS Transfection Kit............................................................................................. 10
TransIT-HeLaMONSTER® Transfection Kit............................................................................. 11
TransIT®-Jurkat Transfection Reagent ................................................................................ 11
TransIT®-Keratinocyte Transfection Reagent ....................................................................... 11
TransIT-Neural® Transfection Reagent................................................................................. 11
TransIT®-Prostate Transfection Kit....................................................................................... 11
LARGE SCALE PROTEIN PRODUCTION
TransIT-PRO® Transfection Kit........................................................................................14-15
3-DIMENSIONAL CELL GROWTH
TransIT®-3D Transfection Reagent/3D Transfection System ................................................ 16
HIGH THROUGHPUT
TransIT®-Express Transfection Reagent............................................................................... 17
OLIGONUCLEOTIDE
TransIT®-Oligo Transfection Reagent................................................................................... 17
PLASMID CONTROLS
Plasmid Delivery Control, Cy™3......................................................................................... 18
Plasmid Delivery Control, Fluorescein.................................................................................. 18
RNA TRANSFECTION
siRNA/miRNA
TransIT-TKO® Transfection Reagent................................................................................19-20
TransIT-siQUEST® Transfection Reagent.........................................................................19-20
LARGE RNA (Viral RNA and mRNA)
TransIT®-mRNA Transfection Kit.....................................................................................21-22
RNAi Controls
RNAi Delivery Control, Cy™3............................................................................................... 23
RNAi Delivery Control, Fluorescein....................................................................................... 23
ELECTROPORATION
PLASMID DNA, RNA, siRNA and miRNA
Ingenio® Electroporation Kit...........................................................................................24-27
Ingenio® Electroporation Solution...................................................................................24-27
Ingenio® Electroporation Accessories.............................................................................24-27
TheTransfectionExperts.com
For Technical Support | Tel +1.608.441.2852 | [email protected]
NEW! TransIT-X2™ DYNAMIC DELIVERY SYSTEM
• Efficient—Exceptional broad spectrum
transfection
PRODUCT NO. QUANTITY
MIR 6003
0.3 ml
• Versatile—Cutting edge delivery of plasmid
DNA and siRNA/miRNA
MIR 6004
0.75 ml
• Technology—Novel, non-liposomal, polymeric
delivery
MIR 6000
1.5 ml
MIR 6005
5 x 1.5 ml
MIR 6006
10 x 1.5 ml
"
DNA/siRNA Transfection
Broad Spectrum DNA & siRNA/miRNA
To inquire about bulk pricing, please call
888.530.0801 (toll free in the U.S.) or
+1.608.441.2852
We recently tested the Trans IT-X2™ Dynamic Delivery System headto-head against Lipofectamine ® 2000 for DNA transfection of NIH-3T3
fibroblasts and the breast cancer cell line ZR-75-1. We observed higher
efficiency and less toxicity when using Trans IT-X2™. We are also pleased
to hear that Trans IT-X2™ will be offered in similar volume configurations
to Lipofectamine ® 2000.
"
Dr. Edwin Li, Assistant Professor
Saint Joseph’s University
Description
Achieve superior transfections with an innovative polymeric system that efficiently delivers both
DNA and RNA out of the endosome and into the cytoplasm, overcoming a critical barrier to nucleic
acid delivery.
TransIT-X2TM Dynamic Delivery System
A549‡
CHO-K1‡
Hep G2‡
HCC 1143‡
HUVEC‡
LNCaP‡
MDA-MB-468‡
MDCK‡
HMEC (epithelial)‡
T47D‡
Immortalized Keratinocytes
FreeStyleTM 293-F
BT-20
NHDF
AU565
NIH-3T3
COS-7
HEK 293
HeLa
PC-3
MCF-7
PC-12
MDA-MB-231
HCC38
RAW 264.7
SK-N-MC
20
6
4
Lipofectamine®
2000
Keratinocytes‡
Caco-2
MDA-MB-453
SH-SY5Y
‡ Cell types with >2-fold luciferase expression in head-to-head comparisons.
FIGURE 1. TransIT-X2™ Dynamic Delivery System Enables Superior Gene Expression in a Variety of Cell Types. TransIT-X2
Dynamic Delivery System and Lipofectamine® 2000 Transfection Reagent were used to transfect plasmid DNA encoding
luciferase into 30 different cell types at three reagent-to-DNA ratios. Luciferase expression was compared at 24 hours
post-transfection using a standard luciferase assay. Head-to-head comparisons at optimized ratios illustrate superior
or equal luciferase expression using TransIT-X2 in 26 of 30 cell types; 11 cell types had expression levels 2-fold higher
than Lipofectamine 2000 (denoted with ‡).
2
DNA/siRNA Transfection >>
TransIT-X2™ Dynamic Delivery System continued
A549
CHO-K1
LNCaP
PC-12
Hep G2
MDCK
HMEC*
NHDF*
-
FIGURE 2. Visualization of High GFP Expression Using TransIT-X2™ Dynamic Delivery System. TransIT-X2 Dynamic
Delivery System was used to transfect plasmid DNA encoding EGFP into A549, CHO-K1, Hep G2, LNCaP, MDCK, PC-12,
primary human mammary epithelial cells (HMEC) and primary normal human dermal fibroblasts (NHDF). Transfections
were performed in 35 mm MatTek dishes using 4-8 μl of TransIT-X2 to deliver 2 μg of DNA. Images (32X) were captured
at 48 hours post-transfection using a Zeiss Axiovert S100 inverted fluorescence microscope. *Indicates primary cell
types.
100
90
EGFP Positive (%)
80
70
60
50
40
30
20
10
0
A549
CHO-K1 Hep G2
MDCK
LNCaP
PC-12
HMEC*
NHDF*
FIGURE 3. High GFP Transfection Efficiency in Multiple Cell Lines and Primary Cells Using TransIT-X2™ Dynamic Delivery
System. TransIT-X2 Dynamic Delivery System was used to transfect plasmid DNA encoding EGFP into A549, CHO-K1,
Hep G2, MDCK, LNCaP, PC-12, primary human mammary epithelial cells (HMEC) and primary normal human dermal
fibroblasts (NHDF). Transfections were performed in 96-well plates using 0.2-0.4 μl of TransIT-X2 to deliver 0.1 μg
of DNA (2:1, 3:1 or 4:1 reagent: DNA ratio). Triplicate wells were assayed 48 hours post-transfection on a guava®
easyCyte™ 5HT Flow Cytometer. *Indicates primary cell types.
3
FIGURE 4. Functional Co-delivery of Plasmid DNA and siRNA
Using TransIT-X2™ Dynamic Delivery System. TransIT-X2
Dynamic Delivery System was used to transfect plasmid
Cy™5 labeled DNA encoding nuclear YFP and Cy™3 labeled
siRNA into HeLa cells. Transfection was performed in a 6-well
plate with Poly-L-Lysine (PLL) coated coverslips using 4 μl
of TransIT-X2 to deliver 2 μg of DNA (2:1 reagent:DNA ratio)
and 25 nM siRNA. Actin cytoskeleton was stained using Alexa
Fluor ® 350 Phalloidin. Image (63X) was captured at 24 hours
post-transfection using a Nikon A1R confocal microscope.
Image key: yellow (nuclear YFP), blue (Cy5 labeled DNA), red
(Cy3 labeled siRNA), green (actin cytoskeleton).
"
"
"
We work on non-small cell lung cancer (NSCLC) which is an adherent cell
culture line. Previously, we have tested many transfection products from
several companies without much success, but the Trans IT-X2™ Dynamic
Delivery System works very well with NSCLC using my protocol.
Dr. Luo Wang,
University of Michigan
Comprehensive Cancer Center
"
"
The Trans IT-X2™ Dynamic Delivery System outperformed all other
transfection reagents we have tested for DNA transfection of our C2C12
mouse myoblast cell line. In addition, Trans IT-X2 was also less toxic.
Dr. G. Du, Assistant Professor
Texas Medical Center
"
We are pleased with the performance of the Trans IT-X2™ Dynamic
Delivery System when transfecting our renal carcinoma cell line
786-0.
"
Sathish Padi,
North Dakota State University
The Trans IT-X2™ Dynamic Delivery System performed
better than our regular transfection reagent (Polyjet)
for delivering DNA into the hard to transfect A549 cell
line. Trans IT-X2 was able to show protein expression
compared to Polyjet which failed to produce detectable
levels of protein containing V5 tag.
"
Jason Liggett and Kyung-Won Min, Baek Lab
University of Tennessee
4
DNA/siRNA Transfection >>
100
90
80
Knockdown (%)
70
60
50
40
30
20
10
0
Transfection Reagent
siRNA Target
TransIT-X2™
Lipofectamine® 2000
TransIT-X2™
aha1
GAPDH
FIGURE 5. TransIT-X2™ Dynamic Delivery System Achieves Higher Knockdown than Lipofectamine® 2000. Human
TransIT-X2 Dynamic Delivery System and Lipofectamine® 2000 Transfection Reagent were used to transfect siRNA
targeting endogenous proteins - GAPDH and aha1 or to deliver a non-targeting control in primary normal human dermal
fibroblasts (NHDF). Cells were transfected in a 6-well plate using 4 μl of TransIT-X2 or 6 μl of Lipofectamine 2000 and
25 nM siRNA according to each manufacturer's protocol. The amount of GAPDH or aha1 mRNA was measured relative to
18s rRNA levels using qRT-PCR and then scaled to the mRNA levels of the negative control, 48 hours post-transfection.
Error bars represent the standard deviation of triplicate wells.
100
Knockdown of PTK9 mRNA (%)
90
80
70
60
50
40
30
20
10
0
miRNA
TransIT-X2™
TransIT-X2™
Pre-miR™ hsa-miR-1
mirVana™ mimic, miR-1
FIGURE 6. Effective miRNA Delivery Using TransIT-X2™ Dynamic Delivery System Yields Decreased Levels of PTK9
mRNA. TransIT-X2 Dynamic Delivery System and Lipofectamine® 2000 Transfection Reagent were used to transfect
T47D cells with Pre-miR™ hsa-miR-1 miRNA Precursor or mirVana™ miRNA mimic, miR-1, both known to decrease
PTK9 mRNA levels. A Pre-miR negative control was transfected to assess baseline mRNA levels. Cells were transfected
in a 12-well plate using 3 μl of TransIT-X2 or Lipofectamine 2000 and 50 nM miRNA according to each manufacturer's
protocol. The amount of PTK9 mRNA was measured relative to 18s rRNA levels using qRT-PCR and then scaled to
the mRNA levels of the negative control, 48 hours post-transfection. Error bars represent the standard deviation of
triplicate wells.
5
Broad Spectrum DNA
TransIT®-2020 TRANSFECTION REAGENT
• Broad Spectrum DNA Delivery—Achieve high
expression in many cell types, including hardto-transfect, insect, stem and primary cells
QUANTITY
MIR 5404
0.4 ml
MIR 5400
1.0 ml
MIR 5405
5 x 1.0 ml
MIR 5406
10 x 1.0 ml
+1.608.441.2852
• Animal Origin Free—provides high
performance with maximum compatibility
IDEAL FOR USE IN VIRUS PRODUCTION
Description
TransIT-2020 Reagent is a versatile transfection solution for broad spectrum DNA delivery
into mammalian cells. This premium reagent is animal component free allowing maximum
compatibility for all downstream applications while outperforming major competitors in most
cell types.
A.
B.
FIGURE 7. TransIT®-2020 Reagent Efficiently Transfects Human Induced Pluripotent Stem (iPS) Cells. TransIT-2020
Transfection Reagent was used to transfect 0.5 x 106 iPS cells with a ZsGreen expressing plasmid (Clontech).
Transfections were performed in 6-well plates using 7.5 µl of TransIT-2020 Transfection Reagent to deliver 2.5 µg of
DNA (3:1, reagent: DNA). Cells were visualized 24 hours post-transfection and imaged at 4X objective with an Olympus
IX71® Inverted Microscope. Image is (A) green fluorescence. Cells were also assayed 24 hours post-transfection on an
Accuri® Cytometer. The histogram (B) shows untransfected cells (black line) compared to cells transfected with plasmid
using TransIT-2020 (green line).
Data courtesy of Cellular Dynamics International.
900,000
70
800,000
60
700,000
50
600,000
500,000
40
400,000
30
300,000
20
200,000
10
100,000
0
2:1
3:1
4:1
TransIT®-2020
6
1.5:1 3:1
5:1
1.5:1:1 3:1:1 5:1:1
Lipofectamine®
2000
Lipofectamine®
LTX
and PLUSTM
0
Toxicity (%)
Luciferase (RLU)
DNA Transfection
• Outperforms Competitor Reagents—
TransIT-2020 demonstrates higher protein
yield and less toxicity when compared to other
transfection reagents
PRODUCT NO. FIGURE 8. TransIT®-2020 Reagent Exhibits Higher
Expression and Lower Cellular Toxicity Compared to Other
Transfection Reagents. Human umbilical vein endothelial
cells (HUVEC) were transfected with a luciferase expression
plasmid using the designated reagents at the reagent-toDNA ratios indicated beneath each bar. Transfections were
performed in 96-well plates using 0.1 µg of plasmid DNA
per well. Luciferase expression (bar graph) and lactate
dehydrogenase (LDH) levels (line graph) were measured
at 24 hours post-transfection. LDH levels are reported
as percent cytotoxicity compared to cells alone and were
measured using a commercially available colorimetric
assay; all values at or below zero are represented as zero
on graph. Error bars represent the standard deviation of
triplicate wells.
DNA Transfection >> TransIT®-2020 Transfection Reagent continued
90
80
70
60
50
40
30
20
10
0
HUVEC
THP-1
TransIT®-2020
NT2
FuGENE® HD
CHO-K1
HEK-293
FIGURE 9. Superior Gene Expression in a Broad Spectrum of Cell Types. The indicated cell types were transfected in
96-well plates with a luciferase expression plasmid (0.1 μg/well) according to industry accepted testing protocols.
Reagent-to-DNA ratios were optimized for each cell type: TransIT®-2020 (Mirus Bio, 2:1 or 3:1), FuGENE® HD (Promega,
3.5:1), Lipofectamine® 2000 (Life Technologies, 1.5:1, 3:1 or 5:1). Luciferase activity was measured 24 hours posttransfection. Values were normalized to TransIT-2020 and presented as a percentage of luciferase expression. FuGENE
is a registered trademark of Fugent LLC. Lipofectamine is a registered trademark of Life Technologies Corporation.
Relative Levels of
Secreted Alkaline Phosphatase
(as determined by O.D. 405 nm)
2.5
2.0
1.5
1.0
0.5
Ef
fec
te
ne
(op
tim
iz
Fe e d 5
cto 0:
Fe Fly 1; 0.
cto (1: 4 u
Fe Fly 1; 0. g)
cto (1: 4 u
1
Fe Fly ( ; 0. g)
ct 1:1 8 u
Fly oFly ; 1. g)
Fe (1: 2 u
Fly ctin 1; 3 g)
Fe (2: .0 u
1
Fly ctin ; 0. g)
F (4 4 u
Fly ecti :1; 0 g)
n ( .4
F
Fu e c 8 :
GE tin 1; 0 ug)
Fu NE (16: .4 u
GE HD 1; g)
0
Fu NE (3:2 .4 u
GE HD ; 0 g)
F NE (5 .4
Ins uGE HD :2; 0 ug)
e
N
( .4
Ins ct G E H 7:2; ug
ec en D ( 0.4 )
e
Ins t Ge Jui 8:2; ug
ec ne ce ( 0.4 )
J
In t Ge uice 5:1; ug
se ne
(1 0. )
ct
G Ju 0 4 u
Lin en ice ( :1; 0 g)
ea eJu 15 .4 u
Lin r P ice :1; g)
ea EI 2 (2 0.4
Lin r PE 5 kD 0:1; ug)
0
ea I 2
5 (5 .4
Lin r PE kD :1; ug)
0.
e I
(
Pr ar P 25 k 10:1 4 ug
om EI D
;0 )
Pr oF 25 (15 .4 u
om ec kD :1; g
t
0 )
Pr oFe in-In (20: .4 u
om cti se 1; g
oF n-In ct ( 0.4 )
Pr ec se 5:
om tin c 1; ug)
oF -In t (1 0.4
ec se 0:1 ug
tin ct ; 0 )
Tr -Ins (15 .4 u
an e :1; g
)
s c
Tr IT-2 t (20 0.4 u
an 0 :1 g
)
;
s 2
Tr IT-2 0 (4 0.4
an 0 :1 ug
Tr sIT 20 ( ; 0.4 )
an -2 6:
sIT 02 1; ug)
-2 0 (8 0.4
02 :1 ug
0( ;0 )
10 .4
:1; ug
0. )
4u
g)
0
Transfection Condition
FIGURE 10. TransIT®-2020 Reagent Effectively Transfects Drosophila S2 Cells. Cells were transfected with a plasmid
construct expressing a secreted form of the Dscam extracellular domain fused to alkaline phosphatase (AP) in a 24well plate. The ratio of transfection reagent-to-DNA and micrograms of DNA per well is noted beneath each bar. All
products were used according to manufacturers’ protocol. All transfections were performed in serum-free media for
4 hours followed by complete media supplementation. An AP enzymatic assay was used to measure the AP levels 24
hours post-transfection.
Data courtesy of a Researcher, University of California, Berkeley.
7
DNA Transfection
Normalized Luciferase Expression (%)
100
Broad Spectrum DNA
TransIT®-LT1 TRANSFECTION REAGENT
• Broad Spectrum DNA Delivery—Utilize one
transfection reagent and protocol for a variety
of cells
• Deliver Single or Multiple Plasmids—
Suitable for many applications such as gene
expression, shRNA expression, virus production
and promoter analysis
"
QUANTITY
MIR 2304
0.4 ml
MIR 2300
1.0 ml
MIR 2305
5 x 1.0 ml
MIR 2306
10 x 1.0 ml
+1.608.441.2852
We routinely use Mirus Trans IT ® -LT1 Transfection Reagent for the delivery of
plasmid DNA to carry out immunoprecipitation experiments. Our lab recently
published using Trans IT ® -LT1 for this application to reveal a crucial
regulator (MCUR1) for calcium uptake in the mitochondria to regulate
cellular metabolism.” (Mallilankaraman, K et al. Nature Cell Biology. December
2012).
"
Dr. Karthik Mallilankaraman,
Madesh Laboratory, Center for Translational Medicine, Temple University
Description
TransIT®-LT1 (Low Toxicity) Reagent is a broad spectrum, high efficiency DNA transfection reagent
that is easy to use and exhibits minimal cellular toxicity. This reagent is a proprietary formulation
of polyamines and cationic lipids that efficiently transfects cells in the presence of serum.
2,500,000
40
35
2,000,000
30
25
1,500,000
20
1,000,000
15
Cytotoxicity (%)
Luciferase (RLU)
DNA Transfection
• Low Cellular Toxicity—Maintain cell density
and reduce experimental biases
PRODUCT NO.
10
500,000
5
0
2:1
Cells
Alone
4:1
3:1
TransIT®-LT1
1.5:1
3:1 5:1 1.5:1 3:1:1 5:1:1 1.5:1 2:5:1 3:5:1
FuGENE® HD
Lipofectamine® 2000 Lipofectamine® LTX
and PLUS™
0
FIGURE 11. TransIT®-LT1 Reagent Exhibits Higher Expression and Lower Cellular Toxicity Compared to Other Transfection
Reagents. Hep G2 cells were transfected with a luciferase expression plasmid using the designated reagents at the
manufacturers' recommended reagent-to-DNA ratio indicated beneath each bar. Transfections were performed in
96-well plates using 0.1 µg of plasmid DNA per well. Luciferase expression (bar graph) and lactate dehydrogenase
(LDH) levels (line graph) were measured at 24 hours post-transfection. LDH levels are reported as percent cytotoxicity
compared to cells alone and were measured using a commercially available colorimetric assay; all values at or below
zero are represented as zero on graph. Experiments were performed as per industry accepted testing protocols. FuGENE
is a registered trademark of Fugent LLC. Lipofectamine is a registered trademark of Life Technologies Corporation.
8
DNA Transfection >> 100
80
60
40
20
-3
PC
H
Ke ep
G
ra
tin 2
oc
yt
es
LN
Ca
N
eu P
ro
-2
a
N
IH
3T
3
3
H
eL
a
7
29
SH
EK
K1
CO
O-
A5
CH
FIGURE 12. TransIT®-LT1 Reagent Efficiently Delivers DNA to a Wide Variety of Cell Lines. Using TransIT-LT1 Transfection
Reagent, cells were transfected with the pEGFP-C1 expression vector, and the percentage of EGFP expressing cells was
determined 24-48 hours post-transfection by flow cytometry.
Luciferase Activity (RLU)
1.0E + 08
1.0E + 07
1.0E + 06
1.0E + 05
CHO-K1
COS-7
HEK 293
TransIT -LT1
®
HeLa
FuGENE 6
®
FIGURE 13. Comparable Luciferase Expression with TransIT®-LT1 Reagent and FuGENE® 6 in Multiple Cell Types. The
indicated cell lines were transfected in duplicate with 1 µg of a luciferase expression vector per well of a 12-well plate
using either 3 µl of the TransIT-LT1 or FuGENE 6 Reagents according to industry accepted testing protocols. Cells were
harvested 24 hours post-transfection and assayed for luciferase activity. FuGENE is a registered trademark of Fugent
LLC.
FIGURE 14. High Efficiency Transfection of iCell®
Cardiomyocytes using TransIT®-LT1 Transfection Reagent.
iCell Cardiomyocytes were plated at 20,000 cells/well in
a 96-well tissue culture plate coated with 0.1% gelatin.
After allowing the cells to recover from thaw, cells were
transfected with 100 ng/well of pMAXGFP (Lonza) using
TransIT-LT1 Transfection Reagent with a 2:1 reagent-toDNA ratio according to the manufacturer’s instructions.
Fluorescent images were taken 3 days post-transfection
using a Olympus IX71® inverted microscope.
9
DNA Transfection
0
49
EGFP Expressing Cells (%)
TransIT®-LT1 Transfection Reagent continued
Cell Line Specific
TransIT® CELL LINE SPECIFIC TRANSFECTION REAGENTS
TransIT® Cell Line Specific DNA Transfection Reagents are formulated to maximize transfection
efficiency while maintaining cellular health in many popular or hard-to-transfect cell types.
All of these reagents offer:
• Optimized Formulations—Designed for each cell type
DNA Transfection
• Low Cellular Toxicity—Maintain cell density and reduce experimental biases due to
toxicity-induced cellular changes
• Serum Compatible—No media changes necessary or extensive optimization required,
saving valuable research time
Product*
Applicable Cell Line(s)
or Cell Type(s)
Efficiency**
Product No.
Quantity
MIR 2704
0.4 ml
MIR 2700
1.0 ml
MIR 2705
5 x 1.0 ml
MIR 2706
10 x 1.0 ml
TransIT®-293 Transfection Reagent
HEK 293,
HEK 293T,
and related
75–85%
To inquire about bulk pricing, please call 888.530.0801 (toll free in the U.S.) or +1.608.441.2852
TransIT -3T3 Transfection Kit (TransIT -3T3 & 3T3 Authority Reagents)
®
®
NIH 3T3, 3T3-L1 and
related
55–65%
MIR 2184
0.4 ml
MIR 2180
MIR 2185
MIR 2186
1.0 ml
5 x 1.0 ml
10 x 1.0 ml
MIR 5504
0.4 ml
MIR 5500
MIR 5505
1.0 ml
5 x 1.0 ml
MIR 5506
10 x 1.0 ml
TransIT®-BrCa Transfection Reagent
MCF-7, MDA-MB-231, MDAMB-453, MDA-MB-468,
T47D
40–80%
TransIT®-CHO Transfection Kit (TransIT®-CHO Reagent & CHO Mojo Reagent)
CHO-K1 and related
50–60%
MIR 2174
0.4 ml
MIR 2170
MIR 2175
MIR 2176
1.0 ml
5 x 1.0 ml
10 x 1.0 ml
TransIT®-COS Transfection Kit (TransIT®-COS Reagent & COS Boss Reagent)
COS-1, COS-7 and related
10
>80%
MIR 2194
0.4 ml
MIR 2190
MIR 2195
MIR 2196
1.0 ml
5 x 1.0 ml
10 x 1.0 ml
DNA Transfection >> Applicable Cell Line(s)
or Cell Type(s)
Product*
Efficiency**
Product No.
Quantity
TransIT-HeLaMONSTER® Transfection Kit (TransIT®-HeLa Reagent and MONSTER Reagent)
50–60%
HeLa and related
MIR 2904
0.4 ml
MIR 2900
MIR 2905
MIR 2906
1.0 ml
5 x 1.0 ml
10 x 1.0 ml
MIR 2124
0.4 ml
MIR 2120
MIR 2125
1.0 ml
5 x 1.0 ml
MIR 2126
10 x 1.0 ml
MIR 2804
0.4 ml
MIR 2800
MIR 2805
1.0 ml
5 x 1.0 ml
MIR 2806
10 x 1.0 ml
MIR 2144
0.4 ml
MIR 2140
MIR 2145
1.0 ml
5 x 1.0 ml
MIR 2146
10 x 1.0 ml
Jurkat, Jurkat-E6, RAW
264.7, THP-1, K562, and
other lymphoid cell lines
10–25%
DNA Transfection
TransIT®-Jurkat Transfection Reagent
TransIT -Keratinocyte Transfection Reagent
®
Immortalized Keratinocyte
20–30%
TransIT-Neural Transfection Reagent
®
C6, Daoy, DB-TRG-05MG,
DI-TNC1, HCN-1A, Neuro-2a,
PC-12, SK-N-MC, SVGp12
>75%
TransIT®-Prostate Transfection Kit (TransIT®-Prostate Reagent and Prostate Boost Reagent)
DU 145, LNCAP, PC-3
>50%
MIR 2134
0.4 ml
MIR 2130
MIR 2135
MIR 2136
1.0 ml
5 x 1.0 ml
10 x 1.0 ml
* Single tube reagents contain the indicated transfection reagent. Transfection reagents with two components are named "Kits" and both components
are listed following the product name.
"
** Transfection efficiency determined by transfection of an EGFP expression vector followed by visual quantification of the percentage of cells expressing EGFP or
via flow cytometry.
Our lab has been satisfied with the routine use of the Trans IT-HelaMONSTER ®
Transfection Kit. Transfections exhibit high target protein expression with
very little cell toxicity. Cells remain viable post-transfection and can be readily
infected with virus without any problems.
"
Dr. Corine St. Gelais,
The Ohio State University − Center for Retrovirus Research
11
Cell Line Specific
• Superior DNA Delivery—Achieve high
expression levels in breast cancer cell types
including: MCF-7, MDA-MB-231, MDA-MB-453,
MDA-MB-468 and T47D
PRODUCT NO.
QUANTITY
MIR 5504
0.4 ml
MIR 5500
1.0 ml
• Formulated for Low Cellular Toxicity—
Maintain cell density and reduce experimental
biases due to toxicity
MIR 5505
5 x 1.0 ml
MIR 5506
10 x 1.0 ml
+1.608.441.2852
Description
TransIT® -BrCa Transfection Reagent is the only breast cancer cell line reagent
available to the market today. While it delivers efficiently to many breast cancer cell
types, it also maintains low cellular toxicity resulting in reduced experimental biases.
90
80
70
EGFP Positive (%)
DNA Transfection
NEW! TransIT®-BrCa TRANSFECTION REAGENT
60
50
40
30
20
10
0
MCF-7
MDA-MB-231 MDA-MB-453 MDA-MB-468
T47D
FIGURE 15. TransIT®-BrCa Transfection Reagent Yields High Efficiency Plasmid DNA Transfection. TransIT-BrCa
Transfection Reagent was used to transfect plasmid DNA encoding GFP into MCF-7, MDA-MB-231, MDA-MB-453,
MDA-MB-468 and T47D breast cancer cell lines. Transfections were performed in 24-well plates using 1-1.5 µl of
TransIT-BrCa Transfection Reagent to deliver 0.5 µg of DNA (2:1 and 3:1, reagent:DNA ratio). Triplicate wells were
assayed 48 hours post-transfection on a guava® easyCyte™ 5HT flow cytometer.
12
DNA Transfection >> TransIT®-BrCa Transfection Reagent continued
20,000,000
40,000,000
MDA-MB-231
MCF-7
15,000,000
Luciferase (RLU)
Luciferase (RLU)
30,000,000
20,000,000
10,000,000
0
2:1
3:1
4:1
TransIT®-BrCa
2:1
3:1
4:1
2:1
10,000,000
5,000,000
0
3:1
4:1
FuGENE® HD
5,000,000
2:1
3:1
4:1
2:1
3:1
4:1
FuGENE® HD
3,000,000
2,000,000
1,000,000
0
3:1
4:1
2:1
TransIT®-BrCa
2:1
3:1
4:1
2:1
3:1
4:1
FuGENE® HD
20,000,000
20,000,000
MDA-MB-468
T47D
Luciferase (RLU)
15,000,000
10,000,000
5,000,000
0
2:1
3:1
4:1
TransIT®-BrCa
2:1
3:1
4:1
2:1
3:1
4:1
FuGENE® HD
10,000,000
5,000,000
0
3:1
4:1
2:1
TransIT®-BrCa
2:1
3:1
4:1
2:1
3:1
4:1
FuGENE® HD
FIGURE 16. TransIT®-BrCa Transfection Reagent Exhibits Higher Luciferase Expression in Breast Cancer Cells Compared to Other Transfection Reagents. Breast Cancer cell lines, MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468
and T47D, were transfected with a luciferase expression plasmid using the designated reagents at the reagent-to-DNA
ratios indicated beneath each bar according to industry accepted protocols. Transfections were performed in 96-well
plates using 0.1 µg of plasmid DNA per well. Luciferase expression was measured at 24 hours post-transfection using
a standard assay. Error bars represent the standard deviation of triplicate wells.
40
Relative Luciferase Units
Luciferase (RLU)
15,000,000
30
20
10
0
E2
0
1nM
10nM
Negative Control
0
1nM
10nM
ERE Reporter Plasmid
MCF-7 (ER positive)
MDA-MB-231 (ER negative)
FIGURE 17. Activation of Estrogen Receptor Pathway is
Detected in MCF-7 Cells Transfected with the TransIT®-BrCa
Transfection Reagent. MCF-7 (estrogen receptor positive)
and MDA-MB-231 (estrogen receptor negative) breast cancer
cells were maintained in complete media containing charcoal
stripped FBS for 3 days prior to transfection. TransIT-BrCa
Transfection Reagent was used to deliver an ERE-luciferase
reporter plasmid or negative control (SA Biosciences) at a
2:1 reagent-to-DNA ratio. Transfections were performed in 96well plates using 0.1 µg of plasmid DNA per well. Twenty-four
hours post-transfection cells were treated with varying levels
of 17-estradiol (E2) for 6 hours. Firefly and Renilla luciferase
expression was measured at 30 hours post-transfection using
a standard assay. Promoter activity is represented as the
ratio of firefly to luciferase relative light units (RLU) using the
Renilla luciferase for normalization. Error bars represent the
standard deviation of triplicate wells.
13
DNA Transfection
MDA-MB-453
4,000,000
Luciferase (RLU)
2:1
3:1
4:1
TransIT®-BrCa
Large Scale Protein Production
• High Performance—Achieve high protein yield
in suspension CHO and 293 cell types
PRODUCT NO.
QUANTITY
MIR 5700
1 ml
• Easy to Use—Compatible with multiple CHO
media formulations
MIR 5760
10 ml
+1.608.441.2852
• Total Cost Savings—Higher protein yield
translates to lower material and labor costs
compared to linear PEI
"
We recently engineered a bispecific immunofusion for the treatment and
elimination of leukemia stem cells. For this work we chose Trans IT-PRO ®
for antibody production of CHO-S cells based on the high protein yield we
obtained.” (Kuo et al, Protein Eng Des Sel. Oct 2012).
"
Jen-Sing Liu, Ph.D.,
Molecular Templates Inc.
Description
Decrease time to produce usable protein by maximizing target protein yields through transient
transfection. The TransIT-PRO® Transfection Kit uses animal origin free components designed for high
and reproducible protein yield in suspension CHO and 293 derived cells. Since it is compatible with
varied media formulations, the same media can be used for both transient and stable expression.
The TransIT-PRO outperforms linear PEI in protein yield, while providing a cost-effective alternative to
FreeStyle™ MAX.
400
A.
Human Fc (µg/L)
DNA Transfection
TransIT-PRO® TRANSFECTION KIT
Day 3
Day 5
Day 7
300
200
100
0
B.
TransIT-PRO® +
25kDa linear PEI
PRO Boost Reagent
—
PRO
PEI
FS
FreeStyle™ MAX
S1
S2
S3
hIgG1
FIGURE 18. Achieve High Antibody Titers Using TransIT-PRO® Transfection Kit in Suspension CHO Cells. IgG1 was
produced by transient transfection using TransIT-PRO and PRO Boost Reagent (1:1:1), 25 kDa linear PEI (6:1) or
FreeStyle™ MAX (1:1) transfection reagents according to the manufacturers' or published protocol (reagent:DNA ratio).
Transfections were performed using 1 µg plasmid DNA per milliliter of culture and 0.5 x 106 cells/ml at the time of
transfection. FreeStyle™ CHO-S cells were cultured in 20 ml of FreeStyle™ CHO Expression medium in 125 ml shake
flasks. (A) Day 3, 5 and 7 supernatants were clarified and analyzed using a human IgG-Fc sandwich ELISA. Error bars
represent the standard deviation of triplicate technical replicates, 25kDa linear PEI is duplicate technical replicates.
(B) Day 7 supernatants were clarified and analyzed by Western blot. An IgG standard was included for quantification
estimate (S1= 1.6 mg/L, S2= 3.2 mg/L, S3 = 6.3 mg/L).
14
DNA Transfection >> TransIT-PRO® Transfection Kit continued
1750
BD SelectTM CD1000 Medium
1500
BD SelectTM CHO Medium
1250
PowerCHO®2 CD Medium
FreeStyleTM CHO Expression Medium
Human Fc (µg/L)
ProCHOTM5 Medium
1000
750
500
250
0
TransIT-PRO® +
PRO Boost Reagent
25kDa Linear PEI
FreeStyleTM Max
12
Yield (mg/L)
10
8
6
4
2
0
1
2
3
4
5
6
7
8
9
10
Protein
TransIT-PRO® Transfection Reagent (Mirus Bio)
293Fectin™ (Life Technologies)
FIGURE 20. Achieve High Protein Yields Using TransIT-PRO® Transfection Kit in Suspension 293 Cells. Ten different
secreted (non‐antibody) proteins were transiently expressed in FreeStyle™ 293‐F cells (Life Technologies) using the
TransIT‐PRO (1.5:1) or 293fectin™ (Life Technologies, 2:1) transfection reagents according to manufacturers protocol.
Cells were grown in FreeStyle™ 293 Expression Medium and transfected at a density of 4 x 106 cells/ml. The scale of the
transfection for each protein varied between 1‐6 L of culture.
Data courtesy of a TransIT‐PRO® pharmaceutical customer.
15
DNA Transfection
FIGURE 19. TransIT-PRO® Provides High Performance Across Varied Media Formulations. FreeStyle™ CHO-S cells were
adapted to five representative growth media as noted in the graph. Cells were transfected with an IgG encoding plasmid
using the TransIT-PRO and PRO Boost Reagent (1:1:1), 25 kDa linear PEI (6:1) (Polysciences), or FreeStyle™ MAX (1:1)
(Life Technologies Corporation) transfection reagents according to published protocol (reagent:DNA ratio). Transfections
were performed in 24-well deep well shaker blocks using 1 µg plasmid DNA per milliliter of culture and 0.5 x 106 cells/
ml at the time of transfection. Human IgG1 was quantitated from day 5 clarified supernatants and analyzed by a human
anti-Fc sandwich ELISA. Error bars represent the standard deviation of triplicate wells.
3-Dimensional Cell Growth
TransIT®-3D TRANSFECTION REAGENT
• High Efficiency DNA Delivery—Achieve high
expression in a wide range of cell types grown
in 3D culture
• Flexibility—Compatible with solid matrix
scaffold and hydrogel 3D culture formats
QUANTITY
MIR 5804
0.4 ml
MIR 5800
1.0 ml
MIR 5805
5 x 1.0 ml
MIR 5806
10 x 1.0 ml
3D Transfection System
Includes: 0.4 ml TransIT-3D Transfection Reagent
and 2 alvetex® 3D 12-well Plates
MIR 5820
each
+1.608.441.2852
Description
Ideal for the transfection of cells in 3D cell culture, the 3D Transfection System combines the
technologies of Reinnervate's alvetex® 3D Cell Culture Plates and Mirus Bio's TransIT®-3D
Transfection Reagent, enabling scientists to create models that more accurately mimic the tissue
environment, gaining a much deeper insight into the complexities of cell function and behavior.
1,500,000,000
1,250,000,000
1,000,000,000
Luciferase Activity (RLUs)
DNA Transfection
• Easy-to-use—User friendly protocol that
requires minimal optimization
PRODUCT NO.
750,000,000
300,000,000
250,000,000
200,000,000
150,000,000
100,000,000
50,000,000
0
Reagent-to-DNA Ratio
2:1
CHO-K1
3:1
HeLa
2:1
HepG2
3:1
MCF-7
2:1
NIH-3T3
FIGURE 21. 3D Transfection of Multiple Cell Types. The indicated cell lines were seeded at optimized cell densities in
12-well alvetex® 3D plates and adapted to 3D culture conditions for 48 hours. Post-adaptation, cells were transfected
with TransIT®-3D combined with a plasmid encoding firefly luciferase at the reagent-to-DNA ratios indicated. Luciferase
activity was measured 24 hours post-transfection using a conventional assay.
FIGURE 22. GFP Expression in 3D Culture. NIH 3T3 fibroblast cells were seeded in an alvetex® 12-well plate and
adapted to 3D growth. Forty-eight hours post-adaptation,
cells were transfected with TransIT®-3D combined with a
plasmid encoding Green Fluorescent Protein (GFP). Cells
were counterstained with the nuclear stain Hoechst 33342
(blue) and visualized via confocal microscopy.
Data courtesy of Reinnervate.
16
DNA Transfection >> High Throughput
TransIT®-EXPRESS TRANSFECTION REAGENT
PRODUCT NO.
QUANTITY
MIR 2004
0.4 ml
• Broad Spectrum DNA Delivery—Utilize one
transfection reagent and protocol for a variety
of cells
MIR 2000
1.0 ml
MIR 2005
5 x 1.0 ml
and reduce experimental biases
MIR 2006
10 x 1.0 ml
+1.608.441.2852
Description
TransIT-Express Transfection Reagent is a broad spectrum, low toxicity DNA transfection reagent
optimized for high-throughput 96-well plate transfections. This reagent is compatible with standard
transfections of adherent cells or increasingly popular reverse transfections.
FIGURE 23. TransIT®-Express Reagent is Designed and
Optimized Specifically for High-throughput and Reverse
Transfections.
Oligonucleotide
TransIT®-OLIGO TRANSFECTION REAGENT
• Unique Formulation—Maximize DNA and RNA
oligonucleotide transfection efficiency in a wide
range of cells
PRODUCT NO.
QUANTITY
MIR 2164
0.4 ml
MIR 2160
1.0 ml
MIR 2165
5 x 1.0 ml
MIR 2166
10 x 1.0 ml
+1.608.441.2852
Oligonucleotides Tested
phosphodiester DNA, phosphothioate DNA (sDNA), phosphothioate RNA (sRNA), 2'OMe RNA,
2'OMe RNA/sDNA Chimerics, and Morpholino/DNA duplexes.
FIGURE 24. TransIT®-Oligo Reagent Achieves High
Transfection Efficiency. HeLa cells were transfected with
Cy™3 and fluorescein labeled phosphothioate DNA oligos
using TransIT-Oligo Reagent and observed 24 hours
post-transfection. The yellow punctate fluorescence is
the result of co-localization of the fluorescein and Cy™3
labeled oligonucleotides after transfection. The image was
acquired using a confocal microscope.
17
DNA Transfection
• High Throughput Reagent—Designed and
optimized specifically for reverse transfections
Plasmid Controls
Label IT® PLASMID DELIVERY CONTROLS
Plasmid Labeling
Label IT® Reagent
Protocol Overview
Chemical
StructureLabeled—Using Mirus established
• Chemically
covalent Label IT® technology
Plasmid
• Sensitive—Directly
Fluorophore track plasmid DNA delivery
or Biotinmicroscopy
usingH fluorescent
Cy™5
N
abel IT Linker
DNA Transfection
®
• Inert and Compatible—Noncoding controls
that areOsuitable for co-delivery experiments
CH3 functional plasmids
with other
N+
CH3
LABEL PRODUCT NO.
QUANTITY
Cy™3
MIR 7904
10 µg
MIR 7905
100 µg
Fluorescein
MIR 7906
Add
Label IT® Reagent
MIR 7907
10 µg
100 µg
• Ready-to-Use—Supplied as 0.5 mg/ml prelabeled plasmid DNA control solution for in vitro
and in vivo tracking studies
Incubate 1 hr @ 37ºC
NH
Description
either Cy™3 or fluorescein labeled 2.7 kb noncoding
Label IT® Plasmid Delivery Controls consist of Purify
plasmid for direct assessment of delivery efficiency in mammalian cells.
N
CH3
Reactive Group
Cl
Covalent Bond
Labeled Plasmids
Transfect Labeled Plasmid and
Detect by Fluorescence Microscopy
Transfect Labeled Plasmid and
FIGURE 25. Ready-to-use Label IT® Plasmid Delivery Controls are Labeled Using the Covalent Label IT® Technology.
Mirus Label IT reagents* are used to chemically attach labels to a noncoding plasmid at optimized label/base pair
ratio to generate Label IT Plasmid Delivery Controls. These pre-labeled Label IT Plasmid Delivery Controls facilitate
direct tracking of plasmid DNA delivery by fluorescence microscopy for in vitro and in vivo studies. The image shows
HeLa cells transfected in serum-containing media with the Label IT Cy™3 Plasmid Delivery Control (red) using the
TransIT®-LT1 Transfection Reagent. Twenty-four hours post-transfection, the cells were fixed, then counterstained to
locate the nuclei (blue) and to stain the actin (green).
*Label IT® kits for a wide variety of applications are also available for purchase.
Visit mirusbio.com/labeling for more information.
18
Express Gene of Interest from Labeled Plasmid
RNA Transfection >> DNA
siRNA/miRNA
TransIT-TKO® & TransIT-siQUEST®
TRANSFECTION REAGENTS
TransIT-TKO® Transfection Reagent
• High Knockdown Efficiency—Achieve
optimal gene silencing in a large percentage
of cells to ensure experimental success
and reduce experimental biases due to
alterations in cellular health
• Flexible Protocol—use with either standard
or reverse transfections
QUANTITY
MIR 2154
0.4 ml
MIR 2150
1.5 ml
MIR 2155
5 x 1.5 ml
MIR 2156
10 x 1.5 ml
TransIT-siQUEST® Transfection Reagent
"
We have tried other transfection reagents, but
only the Trans IT ® -TKO reagent gives us a 100%
transfection rate and gene knockdown without
toxicity in these cells (RAW 264.7).
"
Nature Protocols,
1: 508 - 517 (2006)
PRODUCT NO.
PRODUCT NO.
QUANTITY
MIR 2114
0.4 ml
MIR 2110
1.5 ml
MIR 2115
5 x 1.5 ml
MIR 2116
10 x 1.5 ml
+1.608.441.2852
100
Luciferase Expression (%)
TransIT-siQUEST® Reagent
TransIT-TKO® Reagent
40
30
20
10
o
Ve
r
4.
7
3T
3
W
26
RA
IH
N
CF
-7
M
3
ep
G2
H
29
-7
EK
H
CO
S
Re
a
Al gen
on t
e
CH
OK1
0
FIGURE 26. Knockdown Efficiencies Using TransIT-siQUEST®, TransIT-TKO® Reagents and Lipofectamine® 2000.
Firefly and sea pansy luciferase reporter vectors were co-transfected into various cell lines using TransIT®-LT1
Reagent. Subsequently, firefly luciferase expression was knocked down by transfection of 25 nM anti-firefly luciferase
siRNA using either TransIT-siQUEST (red), TransIT-TKO (tan) or Lipofectamine 2000 (gray) Reagents. Bars indicate
the percent of normalized firefly luciferase expression as compared to each reagent alone control 24 hours posttransfection.
19
RNA Transfection
Description
TransIT-TKO® and TransIT-siQUEST® small interfering RNA (siRNA and miRNA) Transfection
Reagents are broad spectrum reagents that are easy to use and exhibit minimal cellular toxicity.
Each reagent is uniquely formulated and exhibits distinct siRNA/miRNA transfection profiles.
These two reagents allow the user to identify the best transfection reagent for their particular
cell line.
TransIT-TKO® & TransIT-siQUEST® Transfection Reagents continued
Endogenous
Transcript
Cell Line (Source)
100
90
80
70
60
50
40
30
20
10
0
77%
80%
83%
86%
83%
80%
80%
84%
-80%
85%
70%
70%
80%
70%
81%
--
82%
--91%
77%
--82%
92%
-89%
-----82%
TABLE 2. Knockdown of Genes Using
TransIT‑TKO® or TransIT‑siQUEST®
Transfection Reagents. Cells were
transfected with siRNAs targeting
the indicated genes using the
TransIT‑TKO or TransIT‑siQUEST
Reagents, and the knockdown
percentage was determined using
quantitative RT-PCR or luciferase
assays.
*Firefly luciferase expression
vectors were stably integrated
into the parent cell lines and
clonal lines constitutively
expressing firefly luciferase were
used.
non-targeting siRNA luciferase
anti-firefly siRNA luciferase
non-targeting siRNA LDH
anti-firefly siRNA LDH
100
90
80
70
60
50
40
30
20
10
0
Toxicity (%)
Luciferase*
MAPK1
MAPK3
Luciferase*
Luciferase*
Lamin A/C
HeLa (human cervix)
GAPDH
HeLa-luc (human cervix)
Luciferase*
Hepa-luc (mouse liver)
Luciferase*
HepG2 (human liver)
MAPK1
NIH 3T3-lux (mouse fibroblast) Luciferase*
MAPK1
NIH 3T3-L1
MAPK3
Secondary Human Astrocytes
Lamin A/C
ABC A1
Lamin A/C
Primary Mouse Hepatocytes
PPAR-alpha
2.5 µl 3.5 µl
1.5 µl
TransIT-TKO®
0.5 µl
1.0 µl 1.5 µl
Lipofectamine® RNAiMax
0.5 µl
1.0 µl
1.5 µl
0.25 µl 1.25 µl
2.5 µl
DharmaFECT®1
100
90
80
70
60
50
40
30
20
10
0
1.5 µl 2.5 µl
0.5 µl
TransIT-siQUEST®
non-targeting siRNA luciferase
anti-firefly siRNA luciferase
non-targeting siRNA LDH
anti-firefly siRNA LDH
0.5 µl
1.0 µl 1.5 µl
Lipofectamine® RNAiMax
0.5 µl
1.0 µl
1.5 µl
0.25 µl 1.25 µl
2.5 µl
100
90
80
70
60
50
40
30
20
10
0
Toxicity (%)
FIGURE 27. High Efficiency Knockdown and Low Toxicity Using TransIT-TKO® Reagent in HeLa cells. Firefly and
sea pansy luciferase reporter vectors were co-transfected into HeLa cells using TransIT®-LT1 Reagent. Cells were
incubated for at least 4 hours, split into 24-well plates, allowed to adhere and transfected with 25 nM of either a
non-targeting siRNA or an anti-firefly luciferase siRNA using the indicated reagents with the volumes noted beneath
each well. Luciferase expression, normalized to non-targeting siRNA control (bar graph) and lactate dehydrogenase
(LDH) levels (line graph) were measured at 24 hours post-transfection. LDH levels are reported as percent cytotoxicity
compared to cells alone and were measured using a commercially available colorimetric assay; all values at or below
zero are represented as zero on graph.
DNA Transfection
A549-luc (human lung)
BNL CL.2
(mouse liver)
CHO-luc (hamster ovary)
HEK 293-lux (human kidney)
TransIT-TKO® TransIT-siQUEST®
Knockdown
Knockdown
Efficiency
Efficiency
DharmaFECT®1
FIGURE 28. High Knockdown and Low Toxicity Using TransIT-siQUEST® Reagent in CHO Cells Stably Expressing Firefly
Luciferase. CHO-luc cells were grown in 24 wells plates and transfected with 25 nM of either a non-targeting siRNA
or a anti-firefly luciferase siRNA using the indicated reagents with the volumes noted beneath each well. Luciferase
expression, normalized to non-targeting siRNA control (bar graph) and lactate dehydrogenase (LDH) levels (line graph)
were measured at 24 hours post-transfection. LDH levels are reported as percent cytotoxicity compared to cells alone
and were measured using a commercially available colorimetric assay; all values at or below zero are represented as
zero on graph.
20
DNA Transfection >> Large RNA (Viral RNA and mRNA)
TransIT®-mRNA TRANSFECTION KIT
• High Efficiency Delivery—Ensures experimental
success by effectively transfecting RNA into a
large percentage of the cell population
PRODUCT NO.
QUANTITY
MIR 2225
0.4 ml
1.0 ml
MIR 2255
5 x 1 ml
MIR 2256
10 x 1 ml
• Serum Compatible— Perform transfections in the
presence of serum which eliminates the need for a
media change and maintains cellular health
+1.608.441.2852
• Deliver Various Sizes of RNA—Ideal
for specialized applications, such as viral
production, protein expression from mRNA,
and stem cell reprogramming
"
Our lab recently used the Trans IT ® -mRNA Transfection Kit to show that intracellular delivery
of HPLC-purified and pseudouridine-containing mRNA can translate very efficiently without
immune activation which is ideal for mRNA-based gene therapy applications. Trans IT ® -mRNA
further facilitated this work through low toxicity transfections of HEK 293T, human dendritic
cells (DCs) and primary keratinocytes (Karikó et al. Nucleic Acids Research, 39:e142, 2011).
"
Dr. Katalin Kariko, Department of Neurology
University of Pennysylvania
Description
TransIT®-mRNA Transfection Kit provides high efficiency transfection of large RNA molecules
such as mRNA or viral RNA. The kit is easy to use and minimizes cellular toxicity due to its ability
to transfect RNA in the presence of serum.
4,000
% EGFP Positive
Cell Count
75
3,000
50
2,000
25
1,000
Cell Count (viability)
EGFP Positive Cells (%)
100
0
0
TransIT®-mRNA
1:1:1
RNAiMax™
4:1
Stemfect™
2:1
FIGURE 29. High Efficiency and Low Toxicity Transfection Following 14 Consecutive Transfections with TransIT®-mRNA
Transfection Kit. Repeated daily transfections were performed in the same population of BJ fibroblasts using three
commercially available transfection reagents – TransIT-mRNA Transfection Kit (Mirus Bio), Lipofectamine® RNAiMAX
(Life Technologies) and Stemfect™ RNA Transfection Kit (Stemgent) - with a capped and polyadenylated EGFP mRNA
incorporating pseudouridine and 5mC modified bases (Trilink Biotechnologies, Inc.). Multiple reagent-to-RNA ratios
were tested and the optimal ratio is represented. Transfections were performed in 12-well plates using the indicated
reagent-to-RNA ratios to deliver 1 µg of RNA. Transfection efficiency was measured by flow cytometry on a guava®
easyCyte™ 5HT Flow Cytometer following 14 consecutive daily transfections (blue bars). Cell viability was determined
using cell counts measured during flow cytometry (black line grey bars). Error bars represent the standard deviation
of triplicate wells.
21
DNA Transfection
MIR 2250
• Low Cellular Toxicity—Maintain cell density and
reduce transfection induced toxicity
TransIT®-mRNA Transfection Kit continued
Total Luciferase Activity/Well (RLUs)
1,400,000,000
1,200,000,000
1,000,000,000
800,000,000
600,000,000
400,000,000
200,000,000
0
A549
CHO-K1
COS-7
HEK293
HeLa
NIH3T3
Vero
FIGURE 30. High Level Luciferase Expression after Delivery of a Luciferase mRNA using the
TransIT®-mRNA Transfection Kit. Cells in 12-well plates were transfected with a capped and
polyadenylated mRNA encoding luciferase using the TransIT-mRNA Transfection Kit. Approximately 18
hours post-transfection the cells were harvested and the total luciferase activity per well was determined.
90
FIGURE 31. Multiple Dendritic Cell Types Express GFP from
mRNA Transfected using TransIT®-mRNA Transfection Kit.
Murine primary bone marrow derived dendritic cells (BMDC)
and murine dendritic cells types (JAWS II and DC 2.4) were
transfected with 1 μg of capped and polyadenlyated mRNA
encoding GFP using a TransIT-mRNA Reagent: Boost: mRNA
ratio of 1:1:1 (μl:μl:μg). All cells were seeded (80,000 cell/
well) overnight in 24-well plates. Cells were assayed via flow
cytometry 8 hours post transfection. Error bars represent the
standard deviation of at least 3 separate experiments.
80
% GFP+
70
60
RNA Transfection
50
40
30
ary
im
Pr
C
MD
eB
rin
Mu
e
Lin
ell
IC
I
WS
JA
No RNA Control
ine
ll L
Ce
.4
2
DC
Data courtesy of Kyle Phua
(Principal Investigator: Kam W. Leong), Duke University.
MHV RNA Transfected
FIGURE 32. TransIT®-mRNA Transfection Kit Successfully Delivers Viral RNAs 32 kb Long. A 32 kb in vitro transcript
of the murine coronavirus, MHV, was transfected into DBT cells using the TransIT-mRNA Transfection Kit. Successful
transfection assessed by the formation of syncytia 24-48 hours post-transfection. Syncytia were visualized by phase
contrast microscopy.
Data courtesy of Mark Clemenz, Loyola University of Chicago.
22
RNA Transfection >> RNAi Controls
Label IT® RNAi DELIVERY CONTROLS
• Chemically
Labeled—Using Mirus established
LABEL miRNA Labeling
Label IT® Reagent ®
covalent
Label
IT
technology
Protocol OverviewCy™3
Chemical Structure
PRODUCT NO. QUANTITY
MIR 7900
10 µg
• Sensitive—Easily visualize transfected cells
miRNA Sample
MIR 7901
100 µg
and assay delivery efficiency using fluorescent
microscopy CyTM3, CyTM5
Fluorescein
or Biotin
H
Cy™5
MIR 7902
10 µg
• Inert Nand Compatible—Does not target known
MIR 7903
100 µg
mammalian genes or cause off-target effects;
To
inquire
about
bulk
pricing,
please
call
suitable for
O co-delivery experiments with
el IT® Linker
Add 888.530.0801 (toll free in the U.S.) or
CHsiRNA
3
functional
Label IT® Reagent
N+
+1.608.441.2852
CH3
• Ready-to-Use—Supplied as prelabeled siRNA
control 10 µM solution with 10X RNAi Dilution
Buffer
Incubate 1 hr @ 37ºC
NH
Description
Label IT® RNAi Delivery Controls consist of either Cy™3 or fluorescein labeled siRNA duplex
that has the same length, charge and configuration as standard siRNA. The sequence of the
Purify
Label IT RNAi duplex is inert and is not known to affect any cellular events. These controls can
be co-transfected with a functional target gene-specific siRNA and are designed to facilitate
CHsiRNA
assessment
of
delivery efficiency for in vitro and in vivo applications.
3
N
RNA Transfection
Reactive Group
Cl
Covalent Bond
Bond
Covalent
LabelLabeled
IT© RNAi
Delivery
Control
Nucleic
Acid
Hybridize to miRNA-specific Microarray
Slides or Beads
Transfect Label IT RNAi Delivery Control and
FIGURE 33. Ready-to-use Label IT® RNAi Delivery Controls are Labeled Using the Covalent Label IT Technology. Mirus
Label IT reagents* are used to chemically attach labels to a nontargeting siRNA at optimized label/base pair ratio to
generate Label IT Plasmid Delivery Controls. These pre-labeled Label IT RNAi Delivery Controls facilitate direct tracking
of siRNA delivery by fluorescence microscopy for in vitro and in vivo studies. The image shows HeLa cells transfected in
serum-containing media using Label IT Fluorescein RNAi Delivery Control (green) using the TransIT-TKO® Transfection
Reagent. Twenty-four hours post-transfection, the cells were fixed, then counterstained to locate the nuclei (blue) and
the actin (green).
*Label IT® kits for a wide variety of applications are also available for purchase.
Visit mirusbio.com/labeling for more information.
23
Plasmid DNA, RNA, siRNA and miRNA
INGENIO® ELECTROPORATION KITS & SOLUTIONS
• High Efficiency Electroporation—Deliver
DNA or RNA to hard-to-transfect, stem and
primary cells
Ingenio® Electroporation Kits for LonzaAmaxa Nucleofector Devices
• Compatible with Most Conventional
Electroporation Devices—Use your existing
system including Lonza-Amaxa®, Bio-Rad®, or
Harvard BTX®
PRODUCT NO.
QUANTITY
MIR 50112
25 RXN
MIR 50115
50 RXN
• Save Money and Reduce Research Costs
Without Sacrificing Performance—Ingenio
Electroporation Solution is available as a
stand-alone solution or as part of a complete
kit with cuvettes and cell droppers
MIR 50118
100 RXN
(solution, 0.2 cm cuvettes, cell droppers)
Ingenio® Electroporation Kits for All
Other Electroporators, such as Bio-Rad
and Harvard-BTX
(solution, 0.4 cm cuvettes, cell droppers)
Description
Ingenio® Electroporation Solution facilitates
efficient and reliable delivery of nucleic acids to
eukaryotic cells refractory to chemical transfection.
Ingenio is a broad spectrum solution that supports
high efficiency electroporation with minimal toxicity
and replaces standard electroporation solutions
including phosphate buffered saline and serumfree media. Ingenio Kits (include solution,
cuvettes and cell droppers) are compatible
with multiple instruments and facilitate a wide
range of applications requiring nucleic acid
delivery to cells. It is also available as a stand
alone solution.
"
Electroporation
I was very depressed for the last 6 months
because I was unable to transfect my rat cell
line with various transfection reagents. I tried 5
Nucleofection ® programs, 2 buffers and several
different cell densities. But nothing worked. I am
very happy to inform you, Ingenio® is a life saver!
Sanal Madhusudana Girija,
Albert Einstein College of Medicine
24
"
PRODUCT NO.
QUANTITY
MIR 50113
25 RXN
MIR 50116
50 RXN
MIR 50119
100 RXN
Ingenio® Electroporation Solution
PRODUCT NO.
QUANTITY
MIR 50111
25 RXN
MIR 50114
50 RXN
MIR 50117
100 RXN
Ingenio® Electroporation Accessories
Cuvettes
PRODUCT NO.
SIZE
QUANTITY
MIR 50120
0.2 cm 25 PK
MIR 50121
0.2 cm 50 PK
MIR 50122
0.4 cm 25 PK
MIR 50123
0.4 cm 50 PK
Cell Droppers
PRODUCT NO.
QUANTITY
MIR 50124
25 PK
MIR 50125
50 PK
Electroporation >> Ingenio® Electroporation Kits and Solutions continued
100
80
60
40
20
0
HL-60
K562
Jurkat E6-1
SK-N-MC
Ingenio® Solution using Gene Pulser XcellTM System
Ingenio® Solution using Amaxa Nucleofector® 2b Device
Amaxa Nucleofector® Solution V on Amaxa Nucleofector® 2b Device
FIGURE 34. Ingenio® Solution Provides Comparable Efficiency on Amaxa's Nucleofector ® 2b Device. Cells were
electroporated in parallel with an EGFP reporter vector. Two electroporators were used with different electroporation
kits: the Ingenio® Electroporation Kit was used in the Gene Pulser Xcell™ Eukaryotic System (Bio-Rad) and the Amaxa
Nucleofector 2b Device (Lonza); the Amaxa Nucleofector Kit V was used in the Amaxa Nucleofector 2b Device, all
according to manufacturers' recommendations. EGFP expressing cells were identified 24 hours post-electroporation by
flow cytometry and presented as a percentage of the live cell population. Experiments were performed in triplicate on
three separate days and the data averaged.
B.
A.
FIGURE 35. High Efficiency Plasmid DNA Electroporation of Human Induced Pluripotent Stem (iPS) Cells using Ingenio®.
Ingenio Electroporation Kit was used to transfect 2 x 106 iPS cells on the Amaxa® Nucleofector ® 2b Device. Cells were
electroporated with 8 µg ZsGreen expressing plasmid (Clontech) in 100 µl and plated in 6-well plates at 0.33 x 106
cells/well. Cells were visualized 24 hours post-transfection and imaged under 4X objective with an Olympus IX71®
Inverted Microscope. Image is (A) green fluorescence. Cells were also assayed 24 hours post-transfection on an Accuri®
Cytometer. The histogram (B) shows unelectroporated cells (black line) compared to cells electroporated with plasmid
using the Ingenio Electroporation Kit (green line).
"
"
Elizabeth Dominguez,
Cellular Dynamics International (CDI)
25
Electroporation
The Ingenio ® Electroporation Kit is routinely used in our lab to transfect
induced pluripotent stem cells (iPSCs) via electroporation and yields
extremely high transfection efficiency. The Ingenio ® Kit is entirely
compatible with the amaxa Nucleofector ® and is also a much more cost
effective solution.
Ingenio® Electroporation Kits and Solutions continued
100
80
60
40
20
*Ke
ratin
t Co
rtica ocyte
l Ne
uron
*ME
BHK F
-2
CHO 1
-K1
COS
HEK -7
-29
3
HeL
a
Hep
a-SE
A
Hep P
G2
HL60
HU
Jurk VEC
at E
6-1
K56
2
Imm
MCF
orta
7
lize
d K NIH-3T
erat
inoc 3
ytes
PC 12
SKBR3
SKN- M
C
THP
-1
Vero
0
*Ra
*Primary cell types
Nomalized Firefly
FIGURE 36. Ingenio® Electroporation Kits are Ideal for Electroporation in Many Cell Types Using the Amaxa Nucleofector ®
2b Device. Cells were assayed at 24 hours post-electroporation by flow cytometry and reported as percentage of live cell
population. Visit www.TheTransfectionExperts.com for ideal pulse conditions.
100
90
80
70
60
50
40
30
20
10
0
r
r
r
2b
2b
2b
lse
lse
lse
tor
tor
tor
Pu
Pu
Pu
ne
ne
ne
fec
fec
fec
e
e
e
o
o
o
G
e
e
e
G
G
cl
cl
cl
Nu
Nu
Nu
CHO-K1
Electroporation
Luciferase (GL3) siRNA
Jurkat E6-1
HeLa
Non-targeting siRNA Control
FIGURE 37. Ingenio® Kits Effectively Electroporate siRNA in the Amaxa Nucleofector ® 2b and the GenePulser ®
Electroporators. siRNA and plasmid DNA were co-electroporated with the indicated cell lines with the Ingenio
electroporation solution in 0.2 cm cuvettes using either the GenePulser ® (Bio-Rad) or the Amaxa Nucleofector ® 2b
(Lonza). Plasmid encoding firefly luciferase (10 µg/ml ) was co-electroporated with 250nM of either non-targeting
siRNA control or GL3 siRNA into Jurkat E6-1, HeLa and CHO-K1 cells. Twenty-four hours post electroporation, cells were
harvested and assayed for luciferase activity. Data from independent experiments performed on different days were
averaged then scaled to non-targeting siRNA control and are represented as a percentage of the control.
26
Electroporation >> Ingenio® Electroporation Kits and Solutions continued
100
80
60
40
20
K56
2
MC
F- 7
Imo
rtal
NIH
ized
-3T
3
Ker
atin
ocy
tes
PC 12
RAW
264
.7
SHSY5
Y
SKB
R3
SKN-M
C
THP
-1
U93
7
Ver
o
HL60
Jurk
at E
6-1
HeL
a
Hep
G2
te
*ME
F
A54
9-F
lpln
BHK
-21
CHO
-K1
CO
S-7
HEK
-29
3
ocy
*HU
*Ke
ratin
VEC
0
*Primary cell types
FIGURE 38. Ingenio® Electroporation Kits are Ideal for Electroporation in Many Cell Types Using the Bio-Rad®
GenePulser Xcell™ System. EGFP expressing cells were identified 24 hours post-electroporation by flow cytometry and
presented as a percentage of the live cell population. Visit www.TheTransfectionExperts.com for ideal pulse conditions.
A.
B.
100
HL-60
K562
Jurkat E6-1
SK-N-MC
80
60
40
20
0
Ingenio®
Electroporation Solution
PBS
Gene Pulser®
Electroporation Buffer
Propidium Iodide Negative Cells (%)
100
HL-60
K562
Jurkat E6-1
SK-N-MC
80
60
40
20
0
Ingenio®
Electroporation Solution
PBS
Gene Pulser®
Electroporation Buffer
27
Electroporation
FIGURE 39. Ingenio® Kits Outperforms Other Electroporation Solutions in Efficiency and Viability. Cells were
electroporated in parallel with an EGFP reporter vector using either Ingenio Electroporation Solution, PBS or
GenePulser ® Electroporation Buffer (Bio-Rad) on the GenePulser Xcell™ Eukaryotic System. (A) EGFP expressing
cells were identified 24 hours post-electroporation by flow cytometry and presented as a percentage of the live cell
population. (B) Cells were assayed for viablility by propidium iodide staining and flow cytometry analysis. Error bars
represent the standard deviation of triplicate wells.
Terms and Conditions
1. Governing Provisions: All Mirus Bio LLC products
are sold and shipped subject to these Terms and
Conditions, unless modified by a quotation issued by
Mirus Bio LLC or an agreement signed by both Mirus
Bio LLC and the customer. These Terms and Conditions supersede any other written document or oral
discussion including terms or conditions appearing
on a purchase order issued by the customer.
2. Product Safety: Mirus Bio LLC manufactures
products for research use by qualified individuals.
These products may contain chemicals that may be
harmful if misused. Customers are responsible for
reviewing and following the instructions included
in the product protocols and MSDS that accompany
the product. Please contact techsupport@mirusbio.
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Mirus Bio LLC products. We recommend our products
be used in accordance with the NIH guidelines
developed for genetic research.
3. Warranties: Mirus Bio LLC products are warranted
to meet or exceed the stated specifications for the
usable life of the product defined as either the
expiration date for the product or six months from
the date of receipt for those products not carrying an
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has altered or misused the product or failed to store
the product as recommended. The warranty does not
include defects or failures associated with materials
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buyer or any other person to use these products in
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Bio's sole obligation and the customer's sole remedy
are limited to free replacement of products that fail
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THIS WARRANTY IS EXCLUSIVE. MIRUS BIO LLC
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This warranty applies to all products and services
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Unless otherwise agreed, all technical assistance
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make no warranty regarding the accuracy or utility of
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4. Patents: Mirus Bio's products and processes are
covered by one or more patents and are subject to
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Determining the existence and scope of such third
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7. Copyright and Trademarks: All content contained
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Attention: Accounts Payable
Fax: 608.441.2849
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Product that we confirm damaged, incomplete or do
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after shipment or return a product that meets our
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customer must bear the cost of shipping the product
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25% restocking fee.
10. These Terms and Conditions constitute the
entire agreement between Mirus Bio LLC and
Customer with respect to Customer use of the Web
site and Customer purchase of products hereunder,
except as the foregoing (i) may be amended from
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by any express conflicting terms or supplemented by
any express additional terms in a separate written
contract signed by authorized representatives of
Mirus Bio LLC and Customer.
Trademarks
©1996-2013. All rights reserved. Mirus Bio LLC.
All trademarks are the property of their respective
owners. HeLaMONSTER, Ingenio, Label IT, MiraCLEAN,
µArray, pLIVE, siQUEST, TransIT, TransIT-Neural,
TransIT-PRO and TransIT-TKO are registered
trademarks of Mirus Bio LLC. Tracker and TransIT-X2
are trademarks of Mirus Bio LLC. Reagent Agent is a
registered service mark of Mirus Bio LLC.
Other Trademarks
Alvetex is a registered trademark of Reinnervate Limited. FuGENE is a registered trademark of Fugent LLC.
Gene Pulser Xcell is a trademark of Bio-Rad Laboratories, Inc. iCell is a registered trademark of Cellular
Dynamics International. easyCyte is a trademark and
guava is a registered trademark of EMD Millipore.
Lipofectamine is a registered trademark and NCode is
a trademark of Life Technologies Corporation. Amaxa
and Nucleofector are registered trademarks of Lonza
Group Ltd. TransMessenger is a registered trademark
of Qiagen Group.
Registered names, trademarks, etc. used in the Mirus
Bio website, even when not specifically marked as
such, are not to be considered unprotected by law.
External links on this web site are provided only for
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Bio has no interest in, responsibility for, or control
over non-Mirus Bio affiliated linked sites. Mirus Bio
makes no promises or warranties of any kind, express
or implied, including those of merchantability and
fitness for a particular purpose, as to content of
linked sites. In no event shall Mirus Bio be liable for
any damages resulting from use of these links even
if Mirus Bio has been informed of the possibility of
such liability.
Patent and Licensing Information
NOTICE TO PURCHASER: LIMITED LICENSE
Use of certain Mirus Bio TransIT® polyamine
transfection reagents are covered by U.S. Patent
No. 5,744,335, No. 6,180,784, No. 7,101,995, No.
7,601,367 and patents pending. The use of certain
other Mirus Bio transfection products are the subject
of one or more of U.S. Patent No. 7,335,509, No.
7,655,468 and/or other pending U.S. patent applications. Mirus Bio Label IT® nucleic acid labeling
and modifying reagents are covered by U.S. Patent
No. 6,262,252, No. 6,593,465, No. 7,049,142, No.
7,326,780 and No. 7,491,538. Cy™3 and Cy™5
products or portions thereof are manufactured under
license from Carnegie Mellon University and are
covered by U.S. Patent No. 5,268,486. This product is
sold to the Buyer with a limited license for Research
Use Only and is not for clinical, therapeutic or
diagnostic use in humans or animals. This product,
or parts from this product, may not be re-packaged
or re-sold without written permission from Mirus
Bio LLC. The buyer agrees not to infringe upon Mirus
patents or to attempt to reverse engineer, reconstruct,
synthesize or otherwise modify Mirus Bio products. A
license from Mirus Bio LLC is required for commercial
application of this product.
Use of certain TransIT® transfection reagents may be
covered by one or more of US patents No.7,250,479,
No. 7,662,986, No. 7,666,962, and No. 7,714,075
and corresponding claims outside of the US. This
product is sold to the Buyer with a limited license for
the sole purpose of the Buyer using the product as a
transfection agent for in vitro research or evaluation.
This product must not be used in vivo (whether for
therapeutic, diagnostic or clinical use) in humans or
animals. This product must not be used in relation
to any ophthalmic application (including, without
limitation, ophthalmic devices, solutions for ophthalmic use, and devices used to incorporate, deliver
or regulate the release of drugs for the diagnosis,
prophylaxis or treatment of human ophthalmic
diseases). This product (or any part) must not be
re-packaged or resold or otherwise transferred to any
third party without the written permission of Mirus
Bio. The Buyer agrees not to infringe upon Mirus Bio’s
patents or to attempt to reverse engineer, reconstruct,
synthesize or otherwise modify this product. A license
from Mirus Bio may be required for commercial
application of this product, or any use other than in
accordance with the foregoing.
For further information, contact Mirus Bio LLC, 545
Science Drive, Madison WI 53711. Email license@
mirusbio.com.
siRNA
DNA
X2
A Transfection Breakthrough
NEW!TransIT-X2™ Dynamic Delivery System
Achieve superior transfections with an advanced nonliposomal, polymeric system that efficiently delivers
both DNA and RNA out of the endosome and into the
cytoplasm, overcoming a critical barrier to nucleic acid
delivery. The TransIT-X2™ Dynamic Delivery System gives
researchers:
X2
Efficiency–superior
broad spectrum transfection
Delivery–simultaneous
delivery of plasmid DNA and siRNA
X2
X2
Technology–novel,
non-liposomal, polymeric
technology
Functional Co-delivery of Plasmid DNA and siRNA.
See page 4 for experimenal details.
www.TheTransfectionExperts.com
ADVANCE YOUR TRANSFECTIONS.
Request a FREE sample: www.mirusbio.com/samplerequest
Providing gene delivery expertise since 1995
©2013 All rights reserved Mirus Bio LLC. TransIT-X2 is a trademark of Mirus Bio LLC.
PRODUCT LIST
DNA/RNA TRANSFECTION
Broad Spectrum DNA & siRNA/miRNA
Product
NEW! TransIT-X2™
Dynamic Delivery System
Product No. MIR 6003
MIR 6004
MIR 6000
MIR 6005
MIR 6006
Quantity 0.3 ml
0.75 ml
1.5 ml
5 x 1.5 ml
10 x 1.5 ml
Broad Spectrum DNA
MIR 5400
MIR 5405
MIR 5406
MIR 2304
MIR 2300
MIR 2305
MIR 2306
Quantity 0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
Cell Line Specific
Product
TransIT®-293
TransIT®-3T3
Transfection Kit*
NEW! TransIT®-BrCa
TransIT®-CHO
Transfection Kit*
TransIT®-COS Transfection Kit*
TransIT-HeLaMONSTER® Transfection Kit*
TransIT®-Jurkat
TransIT®-Keratinocyte
TransIT-Neural®
TransIT®-Prostate
Transfection Kit*
MIR 2700
MIR 2705
MIR 2706
MIR 2184
MIR 2180
MIR 2185
MIR 2186
MIR 5504
MIR 5500
MIR 5505
MIR 5506
MIR 2174
MIR 2170
MIR 2175
MIR 2176
MIR 2194
MIR 2190
MIR 2195
MIR 2196
MIR 2904
MIR 2900
MIR 2905
MIR 2906
MIR 2124
MIR 2120
MIR 2125
MIR 2126
MIR 2804
MIR 2800
MIR 2805
MIR 2806
MIR 2144
MIR 2140
MIR 2145
MIR 2146
MIR 2134
MIR 2130
MIR 2135
MIR 2136
Quantity 0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
* TransIT® Transfection Kits are supplied with a transfection and a booster reagent.
Large Scale Protein Production
Product
TransIT-PRO®
Transfection Kit
MIR 5760
Quantity 1 ml
10 ml
3-Dimensional Cell Growth
Product
TransIT®-3D
MIR 5800
MIR 5805
MIR 5806
Product
TransIT®-Express
Transfection Kit
MIR 2000
MIR 2005
MIR 2006
Quantity 0.4 ml
1 ml
5 x 1 ml
10 x 1 ml
Oligonucleotide
DNA TRANSFECTION
Product
TransIT®-2020
TransIT®-LT1
High Throughput
Quantity 0.4 ml
1 ml
5 x 1.0 ml
10 x 1.0 ml
Product
TransIT®-Oligo
MIR 2160
MIR 2165
MIR 2166
Quantity 0.4 ml
1.0ml
5 x 1.0 ml
10 x 1.0 ml
Plasmid Controls
Product
Label IT® Plasmid Delivery Control, Cy™3
Label IT® Plasmid Delivery Control, Fluorescein
MIR 7905
MIR 7906
MIR 7907
Quantity 10 μg
100 μg
10 μg
100 μg
RNA TRANSFECTION
siRNA/miRNA
Product TransIT-TKO® Transfection
Reagent
TransIT-siQUEST®
TransIT®-siPAK Kit
MIR 2150
MIR 2155
MIR 2156
MIR 2114
MIR 2110
MIR 2115
MIR 2116
Quantity 0.4 ml
1.5 ml
5 x 1.5 ml
10 x 1.5 ml
0.4 ml
1.5 ml
5 x 1.5 ml
10 x 1.5 ml
0.1 ml of each TransIT-siQUEST® and
TransIT-TKO® Reagents
MIR 2260
each
TransIT®-siPAK Plus Kit
0.1 ml of each TransIT-siQUEST® and TransIT-TKO® Reagents and
10μg Label IT® RNAi Delivery Control, Fluorescein
MIR 2270
each
Large RNA (Viral and mRNA)
Product
TransIT®-mRNA
Transfection Kit
MIR 2250
MIR 2255
MIR 2256
Quantity 0.4 ml
1 ml
5 x 1.0 ml
10 x 1.0 ml
RNAi Controls
Product
Label IT® RNAi Delivery Control, Cy™3
Label IT® RNAi Delivery Control, Fluorescein
MIR 7901
MIR 7902
MIR 7903
ELECTROPORATION
Quantity 10 μg
100 μg
10 μg
100 μg
Product Product No.
Size Ingenio® Electroporation Kits (solution, 0.4 cm cuvettes, cell droppers)
MIR 50113
25 RXN
MIR 50116
50 RXN
MIR 50119
10 RXN
Ingenio® Electroporation Kits (solution, 0.2 cm cuvettes, cell droppers)
MIR 50112
25 RXN
MIR 50115
50 RXN
MIR 50118
10 RXN
®
Ingenio Electroporation
MIR 50111
25 RXN (6.25 ml)
Solution
MIR 50114
50 RXN (12.5 ml)
MIR 50117
10 RXN (25 ml)
Ingenio® Electroporation
MIR 50120
0.2 cm cuvettes (25PK)
Accessories
MIR 50121
MIR 50122
MIR 50123
MIR 50124
Cell Droppers (25 PK)
MIR 50125
Cell Droppers (50 PK)
To inquire about bulk pricing, please call 888.530.0801.

TransIT

Transcription

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