FuGENE® HD Transfection Reagent is a next-generation transfection reagent, free of animal-derived components. It is designed to transfect a wide range of eukaryotic cells including insect cells and many cell lines not transfected well by other reagents （e.g ., MCF-7, RAW 264.7, PC-3, HeLa, MA-10, HepG2, SH-SY5Y, A7r5, STO, SCC-61, STSAR-90, SQ20B, T98）.
Note: For the most up-to-date list of cell types that have been successfully transfected with FuGENE® HD Transfection Reagent, visit our Transfection Special Interest Site at www.powerful-transfection.com and view our Transfection Reference Database. This site also contains much more information about the use of this reagent.
For life science research only.
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Generates meaningful physiological results, since FuGENE® HD Transfection Reagent has low cytotoxicity and doesn't "shock" cells （Figure 1）.
Figure 1: Comparison of transfection efficiency using two transfection reagents under standard medium conditions. After they were transfected with a plasmid carrying LacZ, cells were incubated for an additional 24 hours. β-Galactosidase expression was detected with the β-gal Staining Set from Roche.
Figure 2: Comparison of transfection efficiency with two transfection reagents in the presence of higher serum concentrations. Standard growth medium was replaced by 100% FBS 1 hour prior to transfection. After cells were transfected with a plasmid carrying LacZ, they were incubated for an additional 72 hours. Expression of β-Galactosidase was detected with the β-gal Staining Set from Roche.
- Provides a good model system for mimicking biological conditions, since FuGENE® HD Transfection Reagent can transfect cells in high （up to 100%） serum.
Figure 3: Transfection procedure with FuGENE® HD Transfection Reagent. Please note that the procedure does not require a media change after step 2.
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Generates high levels of protein expression in many adherent and suspension-adapted eukaryotic cell lines , including HEK-293 （Figure 4a）, Hep G2, CHO-S, CHO-K, COS-7, COS-1, and insect cell lines such as High Five （Figure 4b） and Sf9.
Figure 4a: High transfection efficiency and protein expression in suspension-adapted mammalian cells.
Panel a: HEK-293 EBNA suspension-adapted cells were transfected with plasmid DNA expressing GFP. The transfection reagent / DNA ratios used were 7:2, 6:2, 5:2, 4:2, and 3:2.
Panel b: The percentage of cells transfected was determined 28 hours post-transfection, and the quantity of GFP protein was estimated on a Coomassie-stained gel at 72 hours post-transfection.
* estimated yield
Figure 4b: High transfection efficiency and protein expression in High Five insect cells.
Panel a: Cells were transfected with a vector containing the LacZ gene. Transfection efficiency was demonstrated 24 hours post-transfection by histochemical staining for β-galactosidase activity （β-Gal Staining Set）
Panel b: Cells were transfected with vectors expressing human kinases Akt1 and MKK6. Protein expression in cell lysates was measured by western blot at 72 hours post-transfection.
- Saves time and minimizes mistakes, since transfection procedures with
- Simplifies downstream protein purification, since transfections with FuGENE® HD Transfection Reagent often produce high levels of expressed protein.
- Produces protein in insect cells much faster than the baculovirus cell expression system. Protein generating procedures that start with FuGENE® HD Transfection Reagent can take up to 18 fewer days （from transfection to protein purification） than the baclovirus system （Figure 5）.
Figure 5: Comparison of the traditional baculovirus cell expression method with an expression system that uses FuGENE® HD Transfection Reagent.
FuGENE® HD Transfection Reagent take very few steps （Figure 3） and do not need to be customized for different cell types.since FuGENE® HD Transfection Reagent works efficiently in both standard cell culture media （Figure 1） and those that contain up to 100% serum （Figure 2）, eliminating the need for media changes （either before or after transfection）. Composition: unique and proprietary blend of many components, in 80% ethanol.
Storage: stable at +2°C to +8°C.
Stability: 2 years, starting from date of manufacture.
Solution in 80% ethanol, filter-sterilized （0.1 µm filter）, packaged in transparent glass vials.
Lot-specific analysis:
Each lot of FuGENE®HD Transfection Reagent is carefully checked, using ISO DIN 9001-certified procedures, to assure product is performing according to specifications and is consistent from lot to lot. Functional analysis:
Cos 7 cells are transfected in 96 well plates with a reporter gene vector DNA, using 0.3 µl FuGENE® HD and 0.1 µg DNA or 0.25 µl FuGENE® HD and 0.1 µg DNA. Reporter gene activity is monitored via a colorimetric test against a reference lot. Activity must be ≥ 75% compared to a reference lot in at least one of the tested amounts, listed above.
Cytotoxicity analysis:
Cell growth is assessed using the Cell Proliferation Reagent WST-1assay which is an indirect measure of cytotoxicity. The remainder of cells from functional analysis is tested for viability.
