Effectene Transfection Reagent

初代細胞および感受性の高い細胞株へのDNAトランスフェクション

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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

Quantity

1 ml

4 x 1 ml

Inquire

Effectene Transfection Reagentは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

特徴

血清存在下でも高いトランスフェクション効率
少量のDNAで効果的なトランスフェクション
他のトランスフェクション試薬に比べて低い細胞毒性
ハイスループットのスクリーニングに最適

製品詳細

Effectene Transfection Reagentは、様々な細胞株へのDNAトランスフェクション用非リポソーム系脂質の試薬です。Effectene Transfection Reagentは、細胞毒性が非常に低いため初代細胞のような感受性の高い細胞株へのトランスフェクションに最適です。

パフォーマンス

プラスミドDNAのトランスフェクション効率は、Effectene Transfection Reagentが推奨される調製法で用いられる場合、他の試薬よりも高くなります（図" Effectene Reagentによる高トランスフェクション効率"）。Effectene Transfection Reagentは、感度の高い細胞株にオリゴヌクレオチドをトランスフェクションする場合に適しており（図" Effectene Reagentによるオリゴヌクレオチドのトランスフェクション"）、特に初代培養細胞に効果的です（図" 初代細胞における40%のトランスフェクション効率"）。Effectene Reagentによって、多くの細胞株および初代細胞のトランスフェクションが成功しています。細胞型特異的トランスフェクションプロトコールをご用意しています。Effectene Reagentを用いたトランスフェクションは、血清存在下で行なえる上、必要なDNA量も少量（図" 血清、DNA量とトランスフェクション効率の相互関係"）なため、細胞毒性を最小限に抑えることが可能です。

創薬などの分野における組み換えDNAテクノロジーの使用により、ハイスループットトランスフェクションの必要性がますます増加しています。Effectene Reagentを用いたトランスフェクションでは、必要なDNA量が少量で済み、操作手順も簡単です。さらにトランスフェクション複合体を除去する必要がないので、ハイスループットのスクリーニングに最適です。Effectene Transfection Reagentはバルクサイズも購入可能です。

図参照

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は、トランスフェクション効率を高めるために特殊なDNA凝縮エンハンサーと至適化されたバッファーを組み合わせた斬新な非リポソーム系脂質の試薬です。真核細胞へのDNA導入を特に効果的にするため、エンハンサーによりDNA分子を凝縮し、続いてEffectene Reagentの陽イオン脂質でコーティングします。この特長により、トランスフェクション複合体形成の再現性が特に優れています。

操作手順

Effectene製法には2つのステップがあります。まず効率的にDNAを凝縮するために最適な塩濃度のバッファーとEnhancerをDNAとミックスします。インキュベーション時間はわずか2～5分です。その後、Effectene Reagentを添加し、さらに5～10分インキュベートして、Effectene-DNA複合体を形成させます。複合体に培養液（血清および抗生物質を含む）をミックスして、細胞に直接添加します。この細胞は、回収および遺伝子発現解析を行なうまでインキュベートします（フローチャート" Effectene Transfection製法"）。

図参照

Effectene transfection procedure.

アプリケーション

Effectene Transfection Reagentは、幅広い細胞株のトランジェントあるいはステーブルトランスフェクション試薬として最適です。

裏付けデータと数値

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
ApplicationsJA	Plasmid transfection, protein overexpression, reporter studies
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

リソース

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