PolyFect Transfection Reagent

一般的な細胞株への迅速で簡単なDNAトランスフェクション

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PolyFect Transfection Reagent (1 ml)

Cat. No. / ID: 301105

For 25–65 transfections in 60 mm dishes or 50–100 transfections in 6-well plates.

数量

1 ml

4 x 1ml

100 ml

Inquire

PolyFect 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

細胞に特化したプロトコールを使用した至適化されたトランスフェクション
迅速なプロトコールと簡単な操作
高い細胞生存率と低い細胞毒性

製品詳細

PolyFect Transfection ReagentはCOS-7、NIH/3T3、HeLa、293、CHO細胞へのDNAトランスフェクション用に至適化された活性型デンドリマーテクノロジーを利用しています。

パフォーマンス

PolyFect Reagentを用いると、常に高いトランスフェクション効率が得られます（図" CHOおよび293細胞における高いトランスフェクション効率"および" PolyFect Reagentを用いたHeLa細胞へのトランスフェクション"）。PolyFect Transfection Reagentを用いれば、多くのリポソーム試薬とは対照的にトランスフェクション効率が低下することなく血清存在下でのトランスフェクションが可能です。

図参照

High transfection efficiencies in CHO and 293 cells.

PolyFect Reagent with HeLa cells.

原理

PolyFect Transfection Reagentは、特別にデザインされた活性化デンドリマー溶液です。本試薬は、中核から放射状に伸びる分枝を有する既定の球状構造のデンドリマー分子で構成されています（図" 活性化デンドリマー"）。この分子は、プラスに電荷したアミノ基を末端に持ち、これがマイナスに電荷した核酸のリン酸基と相互作用して結合します。PolyFect ReagentはDNAをコンパクトな構造にアセンブルし（図" PolyFect–DNA相互作用"）、それによって細胞表面に結合し、非特異的エンドサイトーシスによって細胞内に取り込まれます。本試薬はエンドソームのpHを緩衝し、エンドソームのヌクレアーゼをpH阻害することによってPolyFect–DNA複合体を安定化させます。

図参照

Activated-dendrimer structure.

PolyFect–DNA interaction.

操作手順

PolyFect Transfection Reagentは即使用可能な溶液です。トランスフェクションの操作手順（フローチャート" PolyFect Transfection製法"）は迅速かつ簡単です。PolyFect ReagentをDNA溶液に添加して混合し、5～10分間のインキュベーション後、培養液（血清および抗生物質を含む）を添加し、PolyFect-DNA複合体をピペットで細胞に添加する。細胞は遺伝子発現のためインキュベートします。複合体の除去あるいは培養液の交換の必要がないので、操作は迅速で簡単です。

ハイスループットトランスフェクション

PolyFect Reagentを用いたトランスフェクションは、操作が簡便で、トランスフェクション複合体を除去する必要もないので、ハイスループットスクリーニングには最適なトランスフェクション試薬です。ハイスループットトランスフェクションにおいて、効率および再現性が非常に高く、細胞毒性も低いPolyFect Reagentは、バルクサイズも用意しております。96ウェルプレートにおけるCOS-7、NIH/3T3、HeLa、HeLa-S3、293、CHO細胞のトランスフェクションの至適化済みのプロトコールについては、QIAGENテクニカルサービスにお問い合わせください。

図参照

PolyFect transfection procedure.

アプリケーション

PolyFect Transfection Reagentは、以下の用途における一般的な細胞株への最適なトランスフェクション試薬です。

遺伝子発現および機能解析
新薬開発および発生分化研究

裏付けデータと数値

High transfection efficiencies in CHO and 293 cells.

Transfection efficiencies for PolyFect Reagent and 2 lipid-based reagents were compared. Cells cultured in 6-well plates were transfected with a beta-galactosidase-reporter plasmid. The appropriate protocol was used for transfection with PolyFect Reagent. Optimized protocols were used for transfection with Reagents L2 and F; these were: 10 µl Reagent L2 with 3 µg DNA and 15 µl Reagent F with 2 µg DNA for CHO cells, and 10 µl Reagent L2 with 2 µg DNA and 7 µl Reagent F with 3 µg DNA for 293 cells. All transfections were performed in triplicate. Two independent transfection experiments are shown for 293 cells.

Specifications

Features	Specifications
Applications	Plasmid transfection, protein overexpression
Transfection type	Transient transfection, stable transfection
Controls	Not included
Features	Optimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum
Technology	Activated dendrimers
Number of possible transfections	Up to 50–100 transfections in 6-well plates / 1 ml reagent
Nucleic acid	DNA
Cell type	COS-7, NIH/3T3, HeLa, 293, CHO cells

リソース

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.

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.

Download Safety Data Sheets for QIAGEN product components.

The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates.

The following protocol is optimized for transient transfection of CHO cells in 96-well plates.

The following protocol is optimized for transient transfection of 293 cells in 96-well plates.

The following protocol is optimized for transient transfection of NIH/3T3 cells in 96-well plates.

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.

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.

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.

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

For optimized transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells

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.

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

For the transfection of oligonucleotides and/or large vector constructs (>20 kb) using PolyFect Transfection Reagent, we recommend performing optimization trials to determine the optimal Transfection Reagent/DNA ratio. The amounts of DNA and Reagent provided in the PolyFect Transfection Reagent Handbook can be used as a guideline.

We suggest to start optimization using a range between half and twice the recommended amount of PolyFect Transfection Reagent while keeping the DNA amount constant, as well as using half and twice the recommended amount of DNA while keeping the amount of PolyFect Transfection Reagent constant. For increasing the transfection efficiency of adherent cells you can use the Attractene Transfection Reagent, or of primary cells and difficult-to-transfect cells using Effectene Transfection Reagent. "

FAQ ID -176

Yes, please follow the Supplementary Protocol 'Transient transfection of HEK 293 cells in 75 cm³ flasks using PolyFect Transfection Reagent' (TFP08).

FAQ ID -1014

Yes, please follow the Supplementary Protocol 'Transient Transfecton of HeLa-S3 cells in 60 mm dishes using PolyFect Transfection Reagent' (TFP01).

FAQ ID -1012

Yes, we have Supplementary Protocols for different cell lines: