Navigation Links
PolyFect Transfection Reagent

Optimized for transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells

Features and benefits
  • Optimized protocols for COS-7, NIH/3T3, HeLa, 293, and CHO cells
  • Fast procedure and easy handling
  • No removal of transfection complexes required
  • Transfection in the presence of serum
  • High efficiency
  • Economical price
Principle

PolyFect Transfection Reagent is a solution of specifically designed activated-dendrimers. The reagent consists of dendrimer molecules of a defined spherical architecture with branches radiating from a central core (1; see figure "Activated-Dendrimer Structure"). The branches terminate at charged amino groups, which can interact with negatively charged phosphate groups of nucleic acids. PolyFect Reagent assembles DNA into compact structures (see figure "PolyFectDNA Interaction"), that bind to the cell surface and are taken into the cell by nonspecific endocytosis. The reagent buffers the pH of the endosome, leading to pH inhibitition of endosomal nucleases, which ensures stability of PolyFectDNA complexes.

Activated-Dendrimer Structure

Schematic representation of an activated dendrimer (1). Note the highly branched structure.

PolyFectDNA Interaction
Model of the SuperFectDNA complex. SuperFect Reagent (purple balls) interacts with DNA (black) to form a ring-like (toroid-like) structure. The upper right section of the illustration shows naked DNA, the lower section shows the interaction between dendrimers and DNA inside the complex, and the upper left section shows the final complete coverage of DNA within the complex.

Procedure

PolyFect Transfection Reagent is provided as a ready-to-use solution. The transfection procedure is fast and simple just add PolyFect Reagent to the DNA solution, mix, incubate for 510 minutes, add growth medium (which can contain serum and antibiotics), and pipet the PolyFectDNA complexes onto the cells. The cells are then incubated for gene expression. No post-transfection removal of complexes or medium change/addition is necessary, making the procedure fast and easy.




Applications


PolyFect Reagent has been specifically developed for fast and easy transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells. Optimized, cell-specific protocols for transfection of these cell lines in different cell-culture formats are provided in the PolyFect Transfection Reagent Handbook. In addition, optimized protocols for transfection of HeLa-S3 cells are available from QIAGEN Technical Services and the Transfection Tools web site (www.qiagen.com/transfectiontools/).

PolyFect Reagent consistently delivers high transfection efficiencies (see figures "High Transfection Efficiencies" and "PolyFect Reagent with HeLa Cells"). In co ntrast to many liposomal reagents, PolyFect Reagent enables transfection in the presence of serum without lowering of transfection efficiency.

High Transfection Efficiencies



Comparison of transfection efficiencies for PolyFect Reagent and two lipid-based reagents. Cells cultured in 6-well plates were transfected with a beta-galactosidasereporter 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 l Reagent L2 with 3 g DNA and 15 l Reagent F with 2 g DNA for CHO cells, and 10 l Reagent L2 with 2 g DNA and 7 l Reagent F with 3 g DNA for 293 cells. All transfections were performed in triplicate. Two independent transfection experiments are shown for 293 cells.

PolyFect Reagent with HeLa Cells



Expression of A -galactosidase and B green fluorescent protein (GFP) in HeLa cells. Cells were cotransfected in 6-well plates with -galactosidase and GFP-reporter plasmids using PolyFect Transfection Reagent and the HeLa cell protocol. Expression was visualized by X-gal staining or fluorescence microscopy 2 days post-transfection.

High-throughput transfection

The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. Transfection using PolyFect Reagent requires minimal handling and removal of transfection complexes is not required, making this reag ent highly suitable for high-throughput screening. PolyFect Reagent provides outstanding transfection efficiencies, excellent reproducibility, and minimal cytotoxicity in highthroughput transfection, and is available in bulk quantities. For optimized protocols for transfection of COS-7, NIH/3T3, HeLa, HeLa-S3, 293, and CHO cells in 96-well plates, please contact QIAGEN Technical Services or visit the Transfection Tools web site at www.qiagen.com/transfectiontools/.

Cited References

1. Tang, M.X., Redemann, C.T., and Szoka, Jr., F.C. (1996) In vitro gene delivery by degraded polyamidoamine dendrimers. Bioconjugate Chem. 7, 703.





'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
2. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
3. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
4. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
5. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
6. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
7. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
8. High-Efficiency Transfections Achieved with New Low-Toxicity Reagent
9. Efficient Transfection of Neurospora Crassa
10. Eppendorf Multiporator Transfection Protocols for Eukaryotic Cells
11. Efficient Transfection of Neurospora Crassa
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:12/2/2016)... , ... December 01, 2016 ... ... will share findings demonstrating the value of DNA microarray comparative genomic hybridization ... year’s San Antonio Breast Cancer Symposium. Using molecular test results from tumors ...
(Date:12/2/2016)... ... December 01, 2016 , ... DrugDev believes the only way ... beautiful technology experience. All three tenets were on display at the 2nd Annual DrugDev ... over 40 sponsor, CRO and site organizations to discuss innovation and the future of ...
(Date:12/2/2016)...   SurePure, Inc. (OTCQB: SURP) a global ... has concluded an agreement with Tamarack Biotics under which ... to acquire units of the Company,s patented photopurification technology ... Concurrently with the option, SurePure has ... will seek regulatory approvals in the United ...
(Date:11/30/2016)... CLEVELAND , Nov. 30, 2016  GenomOncology today announced ... Vice President of Medical Affairs.  Dr. Coleman ... enhancing the company,s proprietary knowledge-enabled platform. The GenomOncology software suite ... of genetic sequencing data and clinical decision support, from quality ... , ...
Breaking Biology Technology:
(Date:12/2/2016)... India , December 1, 2016 ... Authentication type (Fingerprint, Voice), Future Technology (Iris Recognition ... and Region - Global Forecast to 2021", published ... USD 442.7 Million in 2016, and is projected ... at a CAGR of 14.06%.      ...
(Date:12/2/2016)... , Dec. 1, 2016   SoftServe , ... BioLock , an electrocardiogram (ECG) biosensor analysis ... a key IoT asset. The smart system ensures ... vehicle,s steering wheel and mobile devices to easily ... As vehicle technology advances, so too ...
(Date:11/30/2016)... , Nov. 30, 2016 Not many of us realize that we spend ... recovery so we need to do it well. Inadequate sleep levels have been found ... pressure, stroke, diabetes, and even cancer. Maybe now is the best time ... could help them to manage their sleep quality? Continue ... ...
Breaking Biology News(10 mins):