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ROCHE 04709705001- FuGENE® HD Transfection Reagen...

产品信息

 

说明书下载:https://www.roche-applied-science.com
产品规格:1 ml up to 300 transfections
产品货号:
产品名称:FuGENE® HD Transfection Reagent
产品名称4851
Application
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.
Benefits
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.
Prevents cell loss,since FuGENE® HD Transfection Reagent works efficiently in both standard cell culture media (Figure 1) and those that contain up to serum (Figure 2), eliminating the need for media changes (either before or after transfection).
Figure 2: Comparison of transfection efficiency with two transfection reagents in the presence of higher serum concentrations. Standard growth medium was replaced by 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 ) serum.
Saves time and minimizes mistakes, since transfection procedures with FuGENE® HD Transfection Reagent take very few steps (Figure 3) and do not need to be customized for different cell types.
Figure 3: Transfection procedure with FuGENE® HD Transfection Reagent.  Please note that the procedure does not require a media change after step 2.
 
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.

 
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.
 
Product Description
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.
Contents
Solution in 80% ethanol, filter-sterilized (0.1 µm filter), packaged in transparent glass vials.
 
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