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PolyFect Transfection Reagent

快速简单的DNA 转染,用于标准细胞株

  • 优化的细胞特异性转染方案
  • 操作简便迅速
  • 细胞存活率高,毒性低

PolyFect Transfection Reagent采用先进的活化树状多聚物,从多聚物中心向外辐射出带正电荷的氨基基团与DNA 分子结合,为下列细胞株特别优化:COS-7、NIH/3T3、HeLa、293、CHO。

Cat No./ID: 301105
PolyFect Transfection Reagent (1 ml)
For 25–65 transfections in 60 mm dishes or 50–100 transfections in 6-well plates.
Cat No./ID: 301107
PolyFect Transfection Reagent (4 x 1 ml)
For 100–260 transfections in 60 mm dishes or 200–400 transfections in 6-well plates.
Cat No./ID: 301108
PolyFect Transfection Reagent (100 ml)
For 2500–6500 transfections in 60 mm dishes or 5000–10,000 transfections in 6-well plates.

PolyFect Transfection Reagent适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
在CHO和293细胞中具有高转染效率。
对比PolyFect Reagent和两个脂质试剂的转染效率。给培养在6孔孔板中的细胞转染β-半乳糖苷酶报告质粒。使用PolyFect Reagent转染时依据相应实验方案进行。Reagents L2 和F的转染依据优化的实验方案。对于CHO细胞:10 µl Reagent L2转染3 µg DNA,15 µl Reagent F转染2 µg DNA;对于293细胞,10 µl Reagent L2转染2 µg DNA,7 µl Reagent F转染3 µg DNA。所有转染均一式三份进行。图中展示了293细胞的两次独立转染实验。
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PolyFect转染效率。
转染流程简单、快速。将PolyFect Reagent加入DNA溶液中,混匀、孵育5–10分钟,加入培养及(可含有血清和抗生素),将PolyFect-DNA加入细胞中。孵育细胞,进行基因表达。
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活化的树状聚合物结构。
请注意本示意图中的一种活化的树状聚合物的高度支化结构。
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PolyFect–DNA相互作用。
SuperFect-DNA复合物的模型显示了SuperFect Reagent(紫色圆球)与DNA(黑色)相互作用,形成环状结构。图的右上部分显示了裸DNA,下半部分显示了树状聚合物与复合物内部DNA的相互作用,左上部分显示了复合物内DNA的最终完整覆盖。
4
PolyFect Reagent用于HeLa细胞转染。
图中显示了HeLa细胞中[A]β-半乳糖苷酶和[B]绿色荧光蛋白(GFP)的表达。依据PolyFect Transfection Reagent与HeLa细胞转染实验方案,在6孔孔板中将β-半乳糖苷酶和GFP报告质粒共转染入细胞中。转染两天后,通过X-gal染色或荧光显微镜观察基因表达情况。
Performance

PolyFect Transfection Reagent保持高效转染率(参见"High transfection efficiencies in CHO and 293 cells"和"PolyFect Reagent with HeLa cells")。与其他试剂不同的是,PolyFect Transfection Reagent能够在血清存在的情况下进行转染而不降低转染效率。

Principle

PolyFect Transfection Reagent是一种特殊活化的树状聚合物。它由从中心发散出分支的球形大分子构成(参见"Activated dendrimer")。分支终止于带电的氨基,可与带相反电荷的磷酸基相互作用。PolyFect Reagent将DNA聚集成紧密结构(参见 "PolyFect–DNA interaction"),附于细胞表面通过非特异性内吞作用进入细胞。该试剂调节细胞核内pH,抑制核内核酸,从而保证了PolyFect–DNA复合物的稳定性。

Procedure

PolyFect Transfection Reagent是即用型试剂。转染流程(参见"PolyFect transfection procedure")简便迅速,只需要将试剂加入到DNA溶液中混合,反应5–10分钟,加入培养基(可以含血清和抗体),将PolyFect–DNA复合物移到细胞上。培养细胞用于基因表达。无需转染后移除复合物或更换及增加培养基,操作简便快速。

高通量转染

使用PolyFect Transfection Reagent进行转染只需少量手动操作且无需移除转染复合物,特别适用于高通量筛选。PolyFect Reagent在高通量转染中转染效率高,可重复性好,细胞毒性低,适用于批量转染。关于在96孔板中对COS-7、NIH/3T3、HeLa、HeLa-S3、293和CHO细胞进行转染的实验方案,请咨询QIAGEN技术支持。

Applications

PolyFect Transfection Reagent特别适用于标准细胞系高效转染,主要用于:

基因表达与功能研究
药物研究与开发
Features
Specifications
应用 RNAi studies, gene expression studies...
细胞类型 COS-7, NIH/3T3, HeLa, 293, CHO cells
对照 Not included
特点 Optimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum
核酸 DNA
可转染的次数 25–10,000 transfections
技术 Activated dendrimers
转染类型 Transient transfection, stable transfection, plasmid co-transfection

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产品介绍与指南 (1)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
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试剂盒操作手册 (1)
For optimized transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells
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转染实验方案 (2)
Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.
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Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.
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补充实验方案 (13)
The following protocol is optimized for transient transfection of HeLa-S3 cells in 60 mm dishes. Parameters for transfection using other culture formats are given in Table 1. Please note that this protocol is only for HeLa-S3 cells. Optimized protocols for HeLa cells are provided in the PolyFect® Transfection Reagent Handbook.
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The following protocol is optimized for transient transfection of HeLa-S3 cells in 96-well plates. Please note that for HeLa cells the optimal amounts of DNA and PolyFect® Reagent differ from the amounts given in the protocol below. Please call one of the QIAGEN Technical Service Departments or your local distributor for an optimized HeLa protocol.
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The following protocol is optimized for transient transfection of 293 cells in 96-well plates.
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The following protocol is optimized for transient transfection of CHO cells in 96-well plates.
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The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates.
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The following protocol is optimized for transient transfection of HeLa cells in 96-well plates. Please note that for HeLa-S3 cells the optimal amounts of DNA and PolyFect® Reagent differ from the amounts given in the protocol below. Please call one of the QIAGEN Technical Service Departments or your local distributor for an optimized HeLa-S3 protocol.
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The following protocol is optimized for transient transfection of NIH/3T3 cells in 96-well plates.
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用户开发的实验方案 (1)
This procedure has been developed by customers for the transient transfection of HEK 293 cells in 75 cm3 flasks using QIAGEN® PolyFect® Transfection Reagent. It has not been thoroughly tested and optimized by QIAGEN.
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