Description
Based on our proprietary polymer synthesis technology, PolyJet DNA In Vitro
Transfection Reagent is formulated as a biodegradable polymer based DNA transfection reagent that
ensures effective and reproducible transfection on HEK293,
COS-7,
NIH-3T3, HeLa, CHO and a broad ranges of hard-to-transfect mammalian cells. PolyJet reagent is able to
immobilize DNA migration during electrophoresis at very low concentration and
form transfection complex within 5 minutes at RT. A remarkable feature of the
reagent is the rapid and complete degradation of polymer after transfection
complex endocytosis (Figure 1), leading to much less cytotoxicity. PolyJet
reagent, 1.0 ml, is sufficient for ~667 transfections in 24 well plates or ~333
transfections in 6 well plates, providing a very affordable alternative to the
leading products for transfecting a variety of commonly used and
hard-to-transfect mammalian cells.

Figure
1. A Cartoon Showing Biodegradation of PolyJet
DNA Transfection Reagent After Endocytosis of Transfection Complex
Features
- Bio-degradable
after endocytosis
- Exceptional high titers of virus production
- Equally good for very long DNAs (>89 kb)
- Equally good for both single DNA transfection and multi DNA
co-transfection
- High levels of recombinant protein production
- Simple & robust transfection procedure
- Very affordable
Storage
Condition
Store at 4 °C. If stored properly, the product is stable
for 12 months or longer.
|
Cell Lines |
Efficiency (% GFP) |
Cell Lines |
Efficiency (% GFP) |
| McArdle 7777 Hep3D SHEP 3T3-442A COS-7 CV-1 D 407 DHD Pro.b 3LL B16-F10 BAEC BHK-21 Ca Ski CaCo2 CHO HCS-2/8 HEK-293 HeLa HLMEC H-MVEC Huh-7D12 ATT20 SK-N-SH C2C12 HepG2 |
65-70% |
hESCs |
70% |
Examples Showing Transfection Efficiency of PolyJet DNA In Vitro Transfection
Reagent on Commonly Used Cells

Transfection
efficiency comparison of PolyJet vs. lipofectamine Plus on Chinese Hamster
Ovary (CHO) cells. HA tagged beta-tubulin cDNA was delivered into CHO cells
with PolyJet (left panel) and lipofectamine Plus (right panel) respectively.
FITC conjugated antibody against HA tag was utilized to pick up HA-beta-tubulin
(Green) while a DM1a antibody was used to detect endogenous alpha-tubulin
followed by probing with rhodamine conjugated secondary antibody (Red). The
above picture was provided by Dr. Shang Yin of University of Texas at Houston
Medical School as courtesy

A comparison showing
transfection efficiency of PolyJet reagent vs. a leading product, Lipofectamine
2000 on HEK293FT cells. HEK-293FT cells were transfected with GFP vector
(pEGFP-N3) by PolyJet (left panel) and Lipofectamine 2000 (right panel)
respectively. The cells were visualized by Nikon Eclipse Fluorescence microscope
24 hours post transfection

A comparison showing
transfection efficiency of PolyJet reagent vs. a leading product, Lipofectamine
2000 on HepG2 cells. HepG2 cells were transfected with GFP vector (pEGFP-N3)
by PolyJet (left panel) and Lipofectamine 2000 (right panel) respectively. The
cells were visualized by Nikon Eclipse Fluorescence microscope 24 hours post
transfection

Transfection
efficiency comparison of PolyJet vs. Fugene HD on MDCK cells. A
plain GFP DNA was transduced into MDCK cells with PolyJet (left panel) and Fugene
HD (right panel) reagents respectively per manufacturers' protocols.
GFP and DAPI staining were visualized under fluorescence microscopy 48
hours post tansfection. The above comparison data and pictures were
completed and provided by Dr. Ge Zhou of NYU Medical Center as courtesy

A comparison showing
transfection efficiency of PolyJet reagent vs. a leading product, Lipofectamine
2000 on MDCK cells. MDCK cells are notoriously hard to transfect. With
proprietary "Shaved Cell Transfection" protocol, PolyJet (left panel) gives up
to 70% GFP positive cells vs. Lipofectamine 2000 (right panel) around 5%
efficiency. MDCK cells were transfected with GFP vector (pEGFP-N3) by PolyJet
(left panel) and Lipofectamine 2000 (right panel) respectively. The cells were
visualized by Nikon Eclipse Fluorescence microscope 36 hours post transfection

A comparison showing
transfection efficiency of PolyJet reagent vs. a leading product, Fugene HD on
LNCap cells. LNCap cells were transfected with GFP vector (pEGFP-N3) by
PolyJet (left panel) and Fugene HD (right panel) respectively. The cells were
visualized by Nikon Eclipse Fluorescence microscope 24 hours post transfection

A image showing exceptional
transfection efficiency of PolyJet reagent
on Human embryonic stem cells (hESCs).
The
hESCs grown in E8 medium on Geltrexvs
(left panel, DIC imaging) was transfected with pEF1α-GFP.
The GFP expression (right
panel) was visualized by Nikon Eclipse Fluorescence microscope 24 hours post
transfection.
The above pictures were provided by Dr.
Marina Pryzhkova of
Johns Hopkins University as courtesy

Neuro2A cells transfected
with pEGFP-C1 plasmid using PolyJet In Vitro DNA
Transfection Reagent. The Neuro2A cells were visualized by Nikon
Eclipse Fluorescence microscope with DIC phase imaging (left) and FITC
imaging (right) 24 hours post-transfection

Comparison
of cytotoxicity of PolyJet DNA In Vitro Transfection
Reagent with L2K on primary murine skin fibroblast. The primary murine
fibroblast was incubated with the indicated transfection
reagents/pEGFP-C1 (DNA) complexes above for 4 hours in serum-free DMEM
High Glucose medium followed by replacement of complete
serum-containing medium. The cells were visualized by Nikon Eclipse
Fluorescence microscope with DIC phase imaging 24 hours post
transfection
Data Sheet and Protocols
-
General Protocol for Transfecting Mammalian Cells
![]()
-
A Short Protocol for Transfecting Mammalian Cells
![]()
-
Advanced Protocol for Transfecting
Hard-to-Transfect Mammalian Cells
![]()
-
A Protocol for Transfecting Suspension 293 and CHO
Cells
![]()
-
Protocol for Lentivirus Production
![]()
-
Protocol for rAAV Production
![]()
- Technical Note & Transfection Tips
![]()
Great News! We are offering "Wireless PowerPoint Presenter"
to customers who purchase 5x1.0 ml PloyJet or
more by March 2010. Take advantage of it and don't let this chance go...
|
|
|
FG-3: Wireless PowerPoint Presenter with Case for purchase of 5 X 1.0 ml or more. This presenter is offered for limited time only! |
To request a free trial sample, please
Create An Account with us to enter
your shipping address and email us at
order@signagen.com
![]()
Testimonials:
Sorry
for sooo big delay, but now I am totally in love with PolyJet. I have used
PolyJet for a while on mouse ESCs with relatively good efficiency 50~70%.
--------Marina Pryzhkova, Ph.D., JHU
PolyJet Transfection Reagent worked equally as well as lipofectamine
2000, with little evidence of cell death on 293, PC-3 and 22RV1 cells.
I will defiantly consider switching over.
-----Tiffany Wallace, Ph.D., NCI / NIH
I
only did side by side with the testing sample (PolyJet) and Lipo2000
with GFP transfection on COS-7 cells. The result was very good. PolyJet
was even better than L2K.
------Feng Qiao, Ph.D., NEI / NIH
I tested the sample of PolyJet on my NIH-3T3 mouse fibroblasts this
weekend. The results were much better than Lipofecatmine LTX. I'm
attaching a powerpoint slide with my results (I did not quantify the %
transfection efficiency, but the pictures get the point across). I
found that the protocol for difficult-to-transfect cell lines worked
better than the standard protocol.
-----Stephanie Murphy, Ph.D., Dartmouth College
I had chance to try your product finally. It was great success. I used HeLa
cells and got 10% transfection efficiency (<0.1% for Lipofectamine). Thank you!
I was wondering if I also try GenJet Plus DNA In Vitro Transfection Reagent?
According to your website, the reagent works better than regular PolyJet.
-------Yumi Uetake, Ph.D., UMASS
I tested PolyJet and it looks great on MDCK. We placed order. Thank you!
-------Ge Zhou, Ph.D., NYU
We
are happy to provide feedback. PolyJet worked very well for us in HepG2
cells, we got approximately 80% efficiency with pMAX GFP plasmid, by
following the conditions in your suggested protocol. We ran a
comparison with Lipofectamine, which only showed approximately 20-30%
transfection efficiency. We are planning experiments and will be
ordering more soon.
-------Emily Mcallister, PBRC
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
DXY721认为:
悬浮细胞和贴壁细胞在转染过程中差别不大,主要差别在于转染后的筛选,当然如果你做的是瞬时转染就不存在筛选的问题了。
其实转染的过程很简单,问题是能不能转的进去的,转染率能有多少,转进去是否可以稳定表达目的蛋白等等。
我们也是用脂质体做悬浮细胞的转染,说明书上都有具体的操作过程,将脂质体和目的基因按比例混合,然后加到细胞悬液里就OK了,说的简单,实际上还是有一些细节要注意的,比如脂质体和目的基因混合的比例,转染的细胞数,细胞的代数,细胞的状态,有的还要求在转染的前一天传代一次,不过不要怕,这些在脂质体说明书上都有明确的说明,按照说明书做就可以了。
jinghuanlv认为:
悬浮细胞和贴壁细胞转染还是有很大不同的。
脂质体转染的原理基于电荷吸引原理,先形成脂质体-DNA复合物,散布在细胞周围,然后通过细胞的内吞作用,将目的基因导入细胞内,而脂质体复合物与贴壁细胞的接触机会比悬浮细胞高出很多倍,所以,脂质体转染时悬浮细胞的转染效率要明显低于贴壁细胞。
我们实验室转染悬浮细胞是用的电穿孔法,目前为止,悬浮细胞转染的最好方法还是电转,我们实验室用的电转仪是Bio-Rad的,使用条件是电压250V,电容975uF,效果不错,不妨一用。
其次要看下你选择单位的规模如何,上海这边的,你可以看下基尔顿生物,原代细胞培养,动物造模,整体课题外包。
本人研究生
GFP发出绿色荧光的原理是Ca离子进入GFP的beta-barrel结构中引起的特定能级,因此只要这个结构仍然保持着,就可以发出荧光。
由于GFP的beta-barrel结构非常稳定,一些版本的GFP蛋白(如EGFP)甚至能抵抗94C的高温几分钟而不完全变性,因此想在溶液状态下去掉GFP的荧光是很难的,一般需要用光漂白法。
基于其非常稳定的结构,即便细胞被固定了,仍然会有一部分的GFP蛋白保持其构象而发出荧光。此时荧光可能较弱。在荧光显微镜下是有可能看得到的。
各位版友求助,
我使用Hek293构建转染模型,瞬转5质粒,用lipo2000做转染体系。
转染48h,发现荧光较强的细胞都在爬片的边缘,比例十分少。是因为我添加试剂的手法不对吗。
同时也发现加入转染体系后细胞状态特别差。想问一下用lipo2000时可以用无双抗的10%FBSDMEM吗。我转染前6h现在用的是纯DMEM,不含FBS。
请各位大神帮忙
但是有在转染前,将细胞进行重新传代的情况,这样做的目的在于保持细胞的活性状态。
“转染前将细胞以1.5-4.5X104 cells/well 的量(具体接细胞数请参考表1)接种在孔板中,于37℃, 5%CO2 的条件下进行培养,18-24小时( sf9细胞为3-4小时)后转染。”


