LipoExpress™ is specially formulated with proprietary gene delivery enhancers for transfecting HEK293 cells and other mammalian cells. LipoExpress™ offers extremely high transfection efficiencies  for HEK293 related cells as well as many other mammalian cells with less cytotoxicity.  LipoExpress™ (1 ml) is sufficient for 300 to 600 transfections in 24 well plates or 150 to 300 transfections in 6 well plates.

Figure 1. Principle of LipoExpress™ mediated Gene Delivery

Features

- Top choice for hard-to-transfect cells
- Exceptional high titers of virus production
- Suitable for very long DNAs (up to 180 kb)
- High transfection efficiency for  suspension 293 cells (e.g., 293F, 293H, etc)
- High levels of recombinant protein production
- Presence of serum and antibiotics enhances efficiency on 293 cells
- Exceptional efficiency for both single DNA transfection and multi DNAs co-transfection

Figure 2, Comparisons of Transfection Efficiency of LipoExpress™ with Brand Name Products: LipoExpress™ (A),  Lipofectamine 2000 (B) and Fugene HD (C) on HepG2 cells. 

Left Panel: Comparison of transfection efficiency of LipoExpress™ with Lipofecatmine 2000 , and Fugene HD on HepG2 cells. GFP DNA (pEGFP-N3) was transfected with different transfection reagents per manufacturer’s protocols to HepG2 cell (cultured on Collagen pretreated dishes). GFP positive cell (%) and fluorescence intensity were detected by passing through FACS 48 hours post transfection.

Right Panel: Presence of serum and antibiotics enhances LipoExpress™ efficiency on HepG2 cells. HepG2 cell (grown on collagen treated dishes) was transfected with three different conditions: 1, serum and antibiotics free; 2, presence of 10% serum and antibiotics followed by removal 5 hours post transfection and; 3, presence of 10% serum and antibiotics without removal 5 hours post transfection.

Figure 3, Comparison of transfection efficiency of LipoExpress™ (A) with Lipofectamine 2000 (B), TransIT  (C) and Fugene 6 (D) on CHO cells.

Left Panel: Comparison of transfection efficiency of LipoExpress with Lipofecatmine 2000 , TransIT and Fugene 6 on CHO cells. DNAs encoding Renilla luciferase (phRL-CMV) and GFP (pEGFP-N3) were transfected with different DNA transfection reagent per manufacturer’s protocols. Renilla luciferase activity and GFP fluorescence were detected with Renilla Assay System and a Nikon Eclipse fluorescent microcopy respectively 24 hours post transfection.

Right Panel: Comparison of price ($/1.0 ml vial) of LipoExpress™ versus those of Lipofecatmine 2000, TransIT and Fugene 6. All the prices were collected from the manufacturers’ websites

Figure 4, A comparison of transfection efficiency of LipoExpress™ with Lipofectamine 2000  on a hard-to-transfect cell, primary rat aortic smooth muscle cells.

Rat aortic smooth muscle cells were prepared and transfected with pEGFP-N3 by LipoExpress™ (left panel) and Lipofecatmine 2000 (right panel) respectively per manufacturers’ protocols. The transfection efficiency was evaluated by detecting GFP fluorescence with a Nikon Eclipse 2000 microscopy 24 hours post transfection.

Figure 5, A comparison of transfection efficiency of LipoExpress™ (left) with Fugene HD (right) on hard-to-transfect cell, LNCap cells.

The LNCap cells were grown as ATCC recommended procedures and co-transfected with pBabe-hygro-SSeCKs (1.5ug) and pEGFP-N3 (0.5ug) per well (6 well plate) LipoExpress™ (left panel) and Fugene HD (right panel) respectively per manufacturers’ protocols. The transfection efficiency was evaluated by detecting GFP fluorescence with a Nikon Eclipse 2000 microscopy 24 hours post transfection.

Figure 6, Two examples showing exceptional efficiency of LipoExpress™ on hard-to-transfect cells like HepG2 and SaoS-2 cells.

HepG2 and SaoS-2 cells in 95% confluency were transfected with pEGFP-N3 and pSV-β-galactosidase DNAs respectively in presence of serum/antibiotics. The efficiency was checked 48 hours post transfection by Zeiss 510 Confocal Microscopy and β-galactosidase staining kit respectively.

LipoExpress™ DIC	LipoExpress™ FITC
293fectin DIC	293fectin FITC

Figure 7, HEK293 cells transfected with pEGFP-C1 plasmid using LipoExpress™ (upper panel) and Invitrogen 293fectin™ of  (lower panel).

The cells were visualized by Nikon Eclipse Fluorescence microscope with DIC phase imaging (left panel) and FITC imaging (right panel) 24 hours post-transfection.

Figure 8, Comparison of LipoExpress™ and Lipofecatmine LTX (LTX) on generation of Lentivirus (LV).

Three cDNAs were co-transfected with LipoExpress™  and Lipofectamine LTX into 293FT cells. A GFP vector, pHR-SIN-cppt-CMVEWP, was used to determine titer of LV. 1×10⁵ 293F cells per well was plated in to a 24 well plate followed by addition of different of amounts of the vector supernatant, 1 microliter (upper panel) and 10 microliters (lower panel) respectively. 5 days later, the cells was passed with FACS. The numbers at the upper right corner indicate the percentage of transduced cells. The titers of LV generated with LipoExpress™ and LTX were quantified to be 8×10⁶ and 3×10⁶ tu/ml, respectively.

GeneExpresso™ Plus DNA In Vitro Tranfection Reagent is enhanced version of GeneExpresso™ DNA In Vitro Transfection Reagent. GeneExpresso™ Plus is formulated by addition of a booster peptide, giving rise to up to 20 times higher efficiency on variety of mammalian cells in comparison of its previous version. GeneExpresso™ Plus was shown to efficiently deliver genes to various established cell lines as well as primary cells including HEK293, 293T, 293E, CHO, COS1, HeLa, NIH 3T3, insect cell lines (Sf9 and Sf21) and a variety of other eucaryotic cell lines with less toxicity. GeneExpresso™ Plus reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 150 to 300 transfections in 6 well plates.

• Cell-dependent 3~20 times higher efficiency
• Top choice for hard-to-transfect cells
• Equally good for very long DNAs (up to 2.0 mb)
• Efficiency boosted in the presence of serum and antibiotics for most of cell types
• Exceptional high levels of recombinant protein production

Comparisons of Transfection Efficiency of GeneExpresso™ Plus In Vitro DNA Transfection Reagent with the Leading Brand Name Products

GeneExpresso™ Plus is top choice for hard-to-transfect cells. Efficiency comparison of GeneExpresso™ Plus In Vitro DNA Transfection Reagent (left panel) with Lipofectamine 2000 (L2K, right panel) on primary cultured rat aortic smooth muscle cells. A nuclear targeted GFP DNA was transfected with GeneExpresso™ Plus and L2K respectively per manufacturer’s protocols. The efficiency was checked 48 hours post transfection.

Left Panel: Comparison of GeneExpresso™ Plus In Vitro DNA Transfection Reagent with leading products including Lipofectamine 2000 (L2K) and Lipofectamine LTX (LTX) on lentivirus production on 293T cells.
Right panel: Comparison of GeneExpresso™ Plus In Vitro DNA Transfection Reagent with tranfection reagents from other vendors inluding Lipofectamine 2000 (L2K) and FuGene 6 on Hela cells. HeLa cells were transfected with plasmid DNA encoding GFP. Transfections were performed according to manufacturers recommendations and analysed by flow cytometry.

Exceptional high titers of lentivirus generated from 293T cells. Three DNAs co-transfected with GeneExpresso™ Plus In Vitro DNA Transfection Reagent (left panel) in comparison of Lipofectamine LTX (right panel). 1.0 and 10 microliters virus supernatant from 293T cells were used to counter-infected 293F cells which was then passed FACS. The numbers at the upper right corner indicate the percentage of transduced cells. The titers of lentivirus generated with GeneExpresso™ Plus and Lipofectamine LTX were quantified to be 2.1×10⁶ and 9.3×10⁶ tu/ml respectively. 

GeneExpresso is a new transfection reagent. It has the following features:

• High gene delivery efficiency for  difficult-to-transfect cells
• Transfect most mammalian cells
• Extremely low cytotoxicity
• No sera interference
• Very stable
• Simple and quick
• Suitable for high-throughput application…

Broad Transfection Spectrum for Mammalian Cell Types:

Comparisons of GeneExpresso™ In Vitro DNA Transfection Reagent with Popular Brand Products

HEK293 cells transfected with pEGFP-C1 plasmid using GeneExpresso™ in vitro DNA Transfection Reagent (upper panel) and the most popular brand product L2K™ (lower panel). The cells were visualized by Nikon Eclipse Fluorescence microscope with DIC phase imaging (left panel) and FITC imaging (right panel) 24 hours post-transfection

Primary Murine Keratinocytes transfected with pEGFP-C1 plasmid using GeneExpresso™In Vitro DNA Transfection Reagent (upper panel) and one of the most popular brand product FuGene™ 6 (lower panel). The cells were visualized by Nikon Eclipse Fluorescence microscope with DIC phase imaging (left panel) and FITC imaging (right panel) 24 hours post-transfection.