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Signagen/PolyJet™ In Vitro DNA Transfection Reagent/SL100688/0.5 mL_蚂蚁淘,【正品极速】生物医学科研用品轻松购|ebiomall 蚂蚁淘商城
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Signagen/PolyJet™ In Vitro DNA Transfection Reagent/SL100688/0.5 mL
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Signagen/PolyJet™ In Vitro DNA Transfection Reagent/SL100688/0.5 mL
品牌 / 
Signagen
货号 / 
SL100688-5x1.0mL
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4000-520-616
PolyJet™ DNA In Vitro Transfection Reagent

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.

Broad Transfection Spectrum for Mammalian Cell Types

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%
67-76%
68-71%
35%
85-90%
60%
70%
70%
80%
85%
51%
80%
88%
60%
88%
61%
86%
88%
72%
59%
72%
46%
29%
46%
72%

hESCs
SN56

MC3T3-E1
Primary melanocyte
mESCs
L929
MCF-7
MDCK
Neuro2A
NIH 3T3
PC12
SH-SY5Y
SiHa
SKOV3
Huh-7
IGROV1
DF-1, Chicken Embryonic Cell
6CSFMEo
WEHI 231
A549
LNCap
Prim. mouse keratinocyte
Prim. human skin fibroblast
Prim. human pre-adipocyte
Prim. mouse embry. fibroblast

70%
81%
80%
35%
70%
59%
68%
68%
86%
76%
50%
25%
60%
65%
70%
35%
50%
71%
26%
75%
75
29%

50%
32%
30%


Examples Showing Transfection Efficiency of PolyJet™ DNA In Vitro Transfection Reagent on Commonly Used Cells
PolyJet_L2K_CHO
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

PolyJet_L2K_HEK293
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

PolyJet_L2K_HepG2
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

PolyJet_Fugene_HD_MDCK
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

PolyJet_L2K_MDCK
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

PolyJet_Fugene_HD_LNCap
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

PolyJet_hESCs
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

PolyJet_L2K_N2A
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

PolyJet_L2K_Primary_Fibroblast
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 offerin
g "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...

Free Powerpoint presenter

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

Request a free trial sample


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

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细胞转染 123
忻忻相惜4582021-08-01
对于脂质体转染与电穿孔转染都无法成功转染的细胞系建议用病毒感染,此法可以快速100%感染,检测成功率高。向左转|向右转
请教做过DNA 脂质体法转染细胞的高手
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。

HAC15细胞转染,目前用的lip3000、lip2000都用过,不知道是试剂剂量使用问题,还是购买试剂问题,荧光显微镜一个视野下好的有几个荧光,有得没荧光,没荧光,没荧光,求HAC15细胞转染用什么转染试剂转染效率高!!!

细胞转染的详细过程123
dppezmiu2017-10-02
您好,现在这个行业发展的不错,生物实验技术外包也会跟着发展,比如一些高校或者企业部分实验不想自己内部开展,或者涉及的设备比较昂贵,技术要求高,都会寻求外包。但是现在竞争也比较大,的得看单位这边整体做的怎么样。
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。
高效率转染RAW 264.7细胞方法 123
生气的小菊花2016-07-19

本人研究生

最近在Raw264.7细胞上进行干扰和用质粒进行过表达实验,脂质体根本转不进去,后来又换了一个国产的转染试剂转染效率也很低,现在考虑用电转。
我们实验室有Lonza电转仪,有没有好一点的电转染试剂或者细胞转染试剂?

可能有多种原因:目标蛋白对细胞有毒性,导致细胞死亡;转染试剂以及DNA用量信息需要优化,否则对细胞具有伤害;细胞贴壁转染之后没有正常换液。建议: 考虑对目标蛋白进行截短构建、尝试其他细胞系统; 摸索转染试剂以及DNA用量信息,如果转染试剂毒性太大,可以考虑尝试义翘转染试剂sinofection; 对转染后的细胞进行换液处理,如果细胞状态感觉不够理想,可以考虑添加一些血清来帮助细胞恢复健康。以上所有分析、建议的前提是,细胞培养、无菌操作等等都没有问题。祝顺利,加油~
大致是两个因素:转染对细胞有一定损伤,细胞密度大些,有利于减少平均损伤;转染试剂、DNA用量与细胞量有关。
细胞转染GFP质粒并进行表达之后,一般细胞内会有大量的GFP蛋白,可以发出绿色荧光。
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

求助:本人现在做细胞转染,具体实验包括转染目的基因后测MTT,凋亡,侵袭等水平差异,现在有个疑惑,如果MTT结果是促进增殖,那凋亡或侵袭结果的意义是否受其影响,比如同样数量细胞里这个组的凋亡少是否其侵袭能力就强,结果是凋亡少造成的还是有协同作用,如何避免干扰,还有转染后凋亡检测时间的选择,因为不是做的药物而是转染,应如何设置时间,78小时是否有意义?问的有点多,求大家耐心指导

我目前遇到的瞬时转染,没有专门做细胞同步化的。

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。”