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

S_1084_5_GEN_V2

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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.
Copy order details
¥44,500
Quantity
1 ml
4 x 1ml
100 ml
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。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 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

Product Details

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。

Performance

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。

See figures

Principle

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。

See figures

Procedure

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。

See figures

Applications

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

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPlasmid transfection, protein overexpression
Transfection typeTransient transfection, stable transfection
ControlsNot included
FeaturesOptimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum
TechnologyActivated dendrimers
Number of possible transfectionsUp to 50–100 transfections in 6-well plates / 1 ml reagent
Nucleic acidDNA
Cell typeCOS-7, NIH/3T3, HeLa, 293, CHO cells

Resources

Transfection Protocols (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.
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.
MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
Supplementary Protocols (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.
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 CHO cells in 96-well plates.
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 293 cells in 96-well plates.
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.
The following protocol is optimized for transient transfection of NIH/3T3 cells in 96-well plates.
パンフレット (1)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
キットハンドブック (2)
クイックスタートプロトコール (1)
User-Developed Protocols (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.
Certificates of Analysis (1)
Brochures & Guides (1)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
Kit Handbooks (2)
Quick-Start Protocols (1)

FAQ

What are your recommendations for cotransfecting several plasmids?
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
Do you have transfection data for QIAGEN Transfection Reagents?

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
How can I optimize the transfection of oligos and large plasmids using PolyFect Transfection Reagent?

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
Do you have a protocol for transient transfection of HEK 293 cells in 75 cm3 flasks?
FAQ ID -1014
Do you have a protocol for transient transfection of HeLa-S3 cells in 60 mm dishes using PolyFect?

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

FAQ ID -1012
Do you have a protocol for transient transfection of cells in 96-well plates using PolyFect Transfection Reagent?
What is the recipe for 1x PBS solution?

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
Do you have a protocol for transient transfection of Hela-S3 cells using PolyFect Transfection Reagent?

 Yes, we have a Supplementary Protocol for 60 mm dishes and 96-well plates:

FAQ ID -997