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Effectene Transfection Reagent

For DNA transfection of primary cells and sensitive cell lines
  • High efficiency in the presence of serum
  • Efficient transfection with low DNA amounts
  • Far lower cytotoxicity and gentler than many alternatives
  • Suitable for high-throughput screening
Effectene Transfection Reagent is a nonliposomal lipid reagent for DNA transfection into a broad range of cell types. Due to its low cytotoxicity, Effectene Transfection Reagent is highly suitable for transfection of primary cells and many sensitive cell lines.
Cat No./ID: 301425
Effectene Transfection Reagent (1 ml)
€474.00
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1 ml Effectene Reagent, Enhancer, Buffer; for 40 transfections in 60 mm dishes or 160 transfections in 12-well plates
Cat No./ID: 301427
Effectene Transfection Reagent (4 x 1 ml)
€1,501.00
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4 x 1 ml Effectene Reagent, Enhancer, Buffer; for 160 transfections in 60 mm dishes or 640 transfections in 12-well plates
The Effectene Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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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.)
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Serum and DNA quantity vs. transfection efficiency.
The influence of serum and DNA quantity on transfection using Effectene Reagent was examined. COS-7 cells (2 x 104 per well) were seeded in 96-well plates one day before transfection. Cells were transfected using 0.01–1.0 µg beta-galactosidase-reporter plasmid and 0.08–8.0 µl Enhancer (DNA: Enhancer ratio of 1:8) and 2 µl Effectene Reagent, in either the presence or absence of serum. Each bar represents the average efficiency from 4 replicates 48 hours post-transfection.
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Effectene transfection procedure.

DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2-5 minutes. Effectene Reagent is then added and the mixture is incubated for 5-10 minutes to allow Effectene-DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression.

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40% transfection efficiency in primary cells.

Primary rabbit aortic smooth muscle cells were transfected with 0.4 µg of a green fluorescent protein reporter plasmid using Effectene Reagent in 6-well plates. Cells were viewed by fluorescence microscopy 24 hours after transfection. Approximately 40% of the cells were transfected, as determined by FACS analysis. (Data kindly provided by K. Veit, 2nd Medical Clinic, Dept. Clinical Pharmacology, Mainz, Germany.)

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Transfection of oligonucleotides using Effectene Reagent.
C6 glioblastoma cells (1 x 104) were transfected with either naked FITC-labeled CD44 antisense oligodeoxynucleotide (ODN) or with Effectene-ODN complexes. Intracellular accumulation and distribution of the transfected ODNs was analyzed 16 hours post-transfection by immunofluoresence.  [A]  Naked ODNs (2 µg).  [B]  ODNs (0.04 µg) complexed with Effectene Reagent. Transfected ODNs appear yellow in the photos. (Data kindly provided by G. Beutel and M. Schott, Institute of Neuropathology, Hanover Medical School, Hanover, Germany.)
Performance

Higher transfection efficiencies of plasmid DNA are achieved with Effectene Transfection Reagent than with other reagents when used with the recommended procedures (see figure "High transfection efficiencies using Effectene Reagent"). Effectene Transfection Reagent is suitable for transfecting sensitive cell lines with oligonucleotides (see figure "Transfection of oligonucleotides using Effectene Reagent") and is particularly effective for primary cells (see figure "40% transfection efficiency in primary cells"). Many cell lines and primary cells have been successfully transfected using Effectene Reagent; cell type-specific transfection protocols are available. Cytotoxicity is minimal  because transfection with Effectene Reagent can be performed in the presence of serum and requires low amounts of DNA (see figure "Serum and DNA quantity vs. transfection efficiency"). 

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 Effectene Transfection Reagent requires low amounts of DNA and minimal handling. In addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. Effectene Transfection Reagent is available in bulk quantities.

Principle

Effectene Transfection Reagent is an innovative non-liposomal lipid formulation that is used in conjunction with a special DNA-condensing enhancer and optimized buffer to achieve high transfection efficiencies. The enhancer first condenses the DNA molecules and Effectene Reagent subsequently coats them with cationic lipids providing a particularly efficient way of transferring DNA into eukaryotic cells. This feature ensures excellent reproducibility of transfection complex formation.

Procedure

The Effectene procedure has two steps. DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2–5 minutes. Effectene Reagent is then added and the mixture is incubated for 5–10 minutes to allow Effectene–DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression (see flowchart "Effectene transfection procedure").

Applications

Effectene Transfection Reagent is suitable for transient and stable transfection of a broad range of cell types.

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

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Brochures & Guides (2)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
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Supplementary Protocols (7)
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.
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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.
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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.
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