Effectene Transfection Reagent

对原代细胞和敏感细胞系的DNA转染

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Effectene Transfection Reagent (1 ml)

Cat. No. / ID: 301425

1 ml Effectene Reagent, Enhancer, Buffer; for 40 transfections in 60 mm dishes or 160 transfections in 12-well plates

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Quantity

1 ml

4 x 1 ml

Effectene Transfection Reagent 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的（再）订购

特点

在血清存在的情况下进行高效转染
只需使用少量DNA 即可达到高效转染
高转染效率而且较低的DNA使用量
适合高通量筛选

产品详情

Effectene Transfection Reagent是一种非脂质体的脂类转染剂，能将DNA转入多种类型的细胞。因其毒性低，非常适合敏感细胞如原代细胞的转染。

绩效

Effectene Transfection Reagent 的操作流程简单, 转染质粒DNA比用其他的试剂获得的转染效率高（参见" High transfection efficiencies using Effectene Reagent"）。Effectene Transfection Reagent适用于带有寡核苷酸的敏感细胞系的转染（参见" Transfection of oligonucleotides using Effectene Reagent"）。特别适用于原代细胞（参见" 40% transfection efficiency in primary cells"）。使用Effectene Transfection Reagent已成功转染了许多细胞系和原代细胞。有针对不同细胞类型的转染方案。细胞毒性低，因为可在血清存在的情况下使用Effectene Transfection Reagent进行转染且DNA用量低（参见" Serum and DNA quantity vs. transfection efficiency"）。

DNA重组技术应用于药物研发领域，导致对高通量转染需求增加。使用Effectene Transfection Reagent进行转染DNA用量低，手动操作少。此外，无需移除转染复合物，使该试剂高度适用于高通量筛选。Effectene Transfection Reagent适用于批量转染。

查看图表

High transfection efficiencies using Effectene Reagent.

Transfection of oligonucleotides using Effectene Reagent.

40% transfection efficiency in primary cells.

Serum and DNA quantity vs. transfection efficiency.

原理

Effectene Transfection Reagent是一种全新的非脂质体的脂类转染剂，将DNA 与浓缩增强子和优化的缓冲液混合使用以达到高效转染。浓缩增强子先聚合DNA分子，Effectene Transfection Reagent随后用阳离子脂质包围聚合的DNA分子。采用一种特殊有效的方法将DNA转入真核细胞。这种特性保证了复杂复合物转染的可重现性。

程序

Effectene流程分为两步。将DNA与浓缩增强子混合后，加入Effectene Transfection Reagent形成复合物，这一步只需2–5分钟。然后加入Effectene Transfection Reagent，与混合物一起反应5–10分钟以形成Effectene–DNA复合物。复合物与培养基混合（可以含有血清和抗体），直接加入到细胞中。培养细胞至成熟，分析基因表达（参见" Effectene transfection procedure"）。

查看图表

Effectene transfection procedure.

应用

Effectene Transfection Reagent适用于多种类型细胞的瞬时和稳定转染。

辅助数据和图表

High transfection efficiencies using Effectene Reagent.

Transfection efficiencies using Effectene Reagent and two commonly used lipid-based reagents were compared. Murine teratocarcinoma F9 cells (5 x 10⁵) were transfected in 6-well plates with a luciferase-reporter plasmid using optimized conditions based on the manufacturer's instructions for each reagent. Transfection efficiencies were determined by measuring luciferase activity 48 hours post-transfection, and are given as relative light units (RLU). (Data kindly provided by I. Clavereau, D. Petitprez, and I. Van Seuningen, Unité INSERM 377, Place de Verdun, Lille Cedex, France.)

Specifications

Features	Specifications
ApplicationsZH	RNAi studies, gene expression studies, high-throughput transfections
Technology	Non-liposomal lipid formulation in conjonction with a DNA-condensing enhancer
Number of possible transfections	160 transfections in 12-well plates / 1 ml reagent
Cell type	Eukaryotic cells (primary cells and sensitive cells)
Features	Non-liposomal lipid formulation, minimal cytotoxicity
Transfection type	Transient and stable transfection
Controls	Not included
Nucleic acid	DNA

资源

The following protocol is optimized for transient transfection of CHO cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of 293 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The next generation in lipid technology

Brochure detailing reagents for efficient and robust DNA and RNA transfection.

Download Safety Data Sheets for QIAGEN product components.

FAQ

The recommended amount of DNA being transfected should be split between the different plasmids. For example, if you normally transfect 5 µg of one plasmid, use 2.5 µg each for two plasmids (other proportions of the two plasmids can be used, as long as 5 µg total is maintained) as a starting point for optimization.

FAQ ID -124

QIAGEN Transfection Reagents have been used successfully with many different cell types. For your convenience, we have organized data from researchers who have shared their experimental results with us online in our Transfection Cell Database. Simply type your cell line of interest into the 'Search' field on this page, and find transfection results achieved with various QIAGEN Transfection Reagents. Please note that QIAGEN cannot verify data supplied from outside sources.

You can also submit your own transfection data and obtain a gift as appreciation. In addition you can find on our TransFect Protocol Database transfection protocols for specific cell types and plate formats.

FAQ ID -158

Customers have successfully transfected plasmid DNA into insect cell lines S2 and Sf9 using Effectene Transfection Reagent. For a reference for Drosophila S2 cells see QIAGEN News article Issue No. 4, 1999: 'Effectene Reagent yields high transfection efficiencies with Drosophila melanogaster S2 cells'.

Customer data for transfection of DNA into SF9 Spodoptera frugiperda cells using Effectene and SuperFect Transfection Reagent can be found in our Transfection Cell Database.

FAQ ID -397

The composition of 1x PBS solution is:

137 mM NaCl
2.7 mM KCl
4.3 mM Na₂HPO₄
1.47 mM KH₂PO₄

Adjust to a final pH of 7.4.

This solution is not supplied in any QIAGEN Kit, but is used in protocols for various QIAGEN transfection kits.

FAQ ID -1030

Effectene Reagent is a unique non-liposomal lipid formulation. Effectene Reagent is used in conjunction with an Enhancer and a DNA-condensation buffer (Buffer EC) to achieve high transfection efficiencies. In the first step of Effectene–DNA complex formation, the DNA is condensed by interaction with the Enhancer in a defined buffer system. Effectene Reagent is then added to the condensed DNA to produce Effectene–DNA complexes. The Effectene–DNA complexes are mixed with medium and directly added to the cells.

Effectene Reagent spontaneously forms micelle structures that show no size or batch variation, as found with preformulated liposome reagents. This unique feature ensures excellent reproducibility of transfection-complex formation. The process of condensing DNA molecules and then coating them with Effectene Reagent is a particularly effective way to transfer DNA into eukaryotic cells.

Broad cell line spectrum

Effectene Transfection Reagent has been used for transfection of a variety of different cell lines and primary cells, and yields significantly better transfection results than many widely used liposome-based transfection reagents. A searchable list of cell lines and primary cells successfully transfected using Effectene Reagent, as well as customer-developed protocols, is available at the Transfection Tools web site.

FAQ ID -184

The following paramenters can be optimized to increase transfection efficiency when using Effectene Transfection Reagent:

Optimize the Effectene Reagent to DNA ratio

If the ratio of Effectene Reagent to DNA is suboptimal, the overall charge of the complexes may be negative, neutral or strongly positive, which can lead to inefficient adsorption to the cell surface. Optimize the Effectene Reagent to DNA ratio according to the section " Transfection Optimization" of the Effectene Transfection Reagent Handbook.

Increase the amount of Effectene-DNA complex

If the transfection efficiency is lower than expected and cytotoxicity acceptably low, increase the overall amount of Effectene-DNA complexes. See the pipetting scheme in the section "Transfection Optimization" of the Effectene Transfection Reagent Handbook for details.

Optimize the incubation time for gene expression

Different cell types achieve maximal expression levels at different times post-transfection. This should be kept in mind when determining length of incubation after transfection. If the time point of maximal expression is not known for a particular cell line, a time course experiment may be necessary.

Consider vector influences

Factors such as the promoter, origin of replication, and plasmid size influence gene expression rate. The optimal quantity of plasmid DNA used for transfection is dependent on the expression rate of the plasmid.

Optimize cell density at the time of Effectene-DNA complex addition

If cell density is too high at the time of transfection-complex addition, cells may not be at the optimal phase of growth. This can lead to insufficient uptake of the complexes into the cells or insufficient expression of the gene of interest. For adherent cells, the optimal confluency at the time of transfection complex addition is normally 40-80%.

Use high-quality DNA Plasmid DNA used for transfection should be of high quality

Impurities present in the DNA preparation can potentially lower transfection efficiency. DNA should be purified using HiSpeed, QIAfilter, or QIAGEN Plasmid Kits or an equivalent method. For endotoxin-sensitive cell lines and primary cells, we recommend using DNA purified with EndoFree Plasmid Kits to ensure the highest transfection efficiencies.

FAQ ID -181