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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
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Signagen
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SL100688-5x1.0mL
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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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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?

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,做的比较好的,还是上海这边的,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。
个人经验,BHK、CHO是很常用的转染用细胞,而且跟抗体非特异性反应也不高。一般来说,大多数可传代细胞的转染效率都是可以的,还要考虑所用转染剂的类型。

不建议你用MDCK细胞,非常难转染。
细胞转染 实验方法 123
一直攀爬的蜗牛2021-07-31
本人最近正在做质粒DNA的293T细胞转染,请问各位大神,有没有用过sigma的转染试剂,我们实验室买了,但是是凝胶状,不知道怎么用,请指教!
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。
传统的转染试剂有一定的细胞毒性,在转染过程由于提高细胞的通透性因而不能在培养基中添加抗生素。如,阳离子脂质体。带正电的脂质体与核酸带负电的 磷酸基团 形成复合物被细胞内吞。血清中含有大量的蛋白质,在转染过程中,带负电的蛋白质可能干扰阳离子脂质体对核酸的吸附,影响转染效率。另外,使用脂质体等转染 试剂时,由于含血清转染会将血清中的蛋白带入细胞,引发细胞毒性,导致转染效率降低,故不能有血清转染。这些转染试剂要求在转染前换无血清培养基,转染后 4-6h在更换成有血清培养基,这其实是为了减轻转染造成的细胞毒性。

我刚开始做转染,悬浮细胞,分别做过表达和敲减,看了很多文献,大都没有提及转染后是用转染的这同一批细胞同时做pcr,wb,cck8,凋亡,细胞周期;还是说这次转染只做pcr或wb,再转染一次做cck8或细胞周期。剩下的功能试验均同前,转染一次做一次?我养的是悬浮细胞,转染后做cck8这些功能试验前需要离心换液吗?跪谢解答!

转染6孔板后,一般长到十的六次方级别就可以提蛋白了
可以用移液器将细胞吸出来并高速离心,沉淀重悬于PBS中洗涤,接着就可以裂解提取蛋白了。可以用超声,酶解等等,裂解后离心收集上清。
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。

本人为新手,希望大家能够直到一下细胞转染效率的问题。现在已知的信息如下:

质粒上面包含了萤光虫荧光素酶基因和海参荧光素酶基因,请问我通过什么方法可以测得转染效率?

如果不能测得转染效率,请问怎么优化转染条件呢?

质粒转染细胞后多久能提RNA做PCR
实验室一直都是用日常型质粒抽提试剂盒,转染细胞没问题。
我觉得只要是注意以下2点就可以了:
1,注意大肠杆菌(Escherichia coli)本身的污染,收集菌体沉淀时防止菌液散落,经常用75%的乙醇擦拭手套。
2,最后洗脱时最好使用无内毒素的水,我们是用注射用水的。
无论是小提还是大提我们都是用的日常型的,并没有刻意用转染级的,因为转染量大,去内毒素的操作太麻烦,损失太大。
可以传代。
转染分2种,一种是瞬时转染,即转染后让细胞表达目的蛋白后即提取蛋白,提一次蛋白,转染一次,这种方式一般不传代;
另一种转染为稳定转染,转染后加入一定选择压力进行筛选,没有转染的细胞不能存活,只留下转染的细胞,这种情况下可以筛选单个转染细胞,构建稳定表达某一特定蛋白或基因的细胞系。