Lipofectamine® 3000 Reagent

Superior performance into the broadest spectrum of difficult-to-transfect cell types

Lipofectamine® 3000 reagent leverages our most advanced lipid nanoparticle technologies to enable superior transfection performance and reproducible results. It delivers exceptional transfection efficiency into the widest range of difficult-to-transfect and common cell types (Figure 1) with improved cell viability.



Figure 1. Lipofectamine® 3000 reagent outperforms Lipofectamine® 2000 and FuGENE® HD reagents. Each reagent was used to transfect HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines in a 96-well format, and GFP expression was analyzed 48 hr posttransfection. Lipofectamine® 3000 reagent provided higher GFP transfection efficiency than Lipofectamine® 2000 and FuGENE® HD reagents for all five cell lines.

Figure 2. Western blot showing GST-STAT protein expression in HepG2 cell line. Cells were transfected using Lipofectamine® 2000 reagent, Lipofectamine® 3000 reagent, FuGENE® HD reagent (competitor 1), and X-tremeGENE® HP reagent (competitor 2) with GST-STAT plasmid in a 24-well plate. 48 hr posttransfection, cells were harvested and western blot analysis was performed. β-actin blot shows all four samples were equally loaded.

Improved cell viability, low toxicity

When developing Lipofectamine® 3000 we optimized all four steps in the transfection process and combined our most advanced lipid nanoparticle technologies together. The superior transfection performance allows you to reduce the reagent dose and lower any potential toxicity risk when working with your cell line.

Figure 3. Each reagent was used to transfect HeLa cells in a 96-well format at the indicated doses with an emerald green fluorescent protein (GFP)–expressing vector. Analysis was performed 48-hours posttransfection utilizing flow cytometry to determine percent transfection efficiency and the intensity of GFP expression. Lipofectamine® 3000 reagent delivered higher efficiency and protein expression than both Lipofectamine® 2000 reagent and Lipofectamine® LTX reagent.

Table 1. Lipofectamine® 3000 reagent yields higher transfection efficiencies than Lipofectamine® 2000 reagent when tested in a variety of cell lines.

Enhance your research in cancer

Using the most biologically relevant cell lines provides the most relevant answers to your research questions. The superior cell spectrum and higher transfection efficiency of Lipofectamine® 3000 reagent enables excellent flexibility to use your cell line of choice in your studies. Lipofectamine® 3000 reagent demonstrates significantly higher transfection performance compared to leading transfection reagents such as Lipofectamine® 2000 reagent in a panel of cancer cells derived from various tissues. While only 25% of the selected cancer cell line panel would be considered easy-to-transfect (>50% transfection efficiency) with Lipofectamine® 2000 reagent, 60% of the panel is easy-to-transfect with Lipofectamine® 3000 reagent.

Table 2. Lipofectamine® 3000 reagent yields higher transfection efficiencies than Lipofectamine® 2000 reagent when tested in cancer cell lines.

Cancer cell line	Lipofectamine® 3000 reagent transfection efficiency	Fold protein expression improvement, Lipofectamine® 3000 vs 2000 reagent
Hs 578.T		3
SW480		2
SK-MEL-28		2
4T1		2
RD		4
Saos-2		4
PANC-1		3
NCI-H23		2
BT-549		4
L6		8
SK-OV-3		3
DU 145		2
HCC1937		5
SW620		5
MDA-MB-468		9
NCI-H460		17
Caki-1		4

Transfection efficiency (%): <30%     30–50%    51–79%    >80%

Figure 4. Lipofectamine® 2000 reagent and Lipofectamine® 3000 reagent were used to transfect 17 cell lines with a GFP-expressing plasmid in a 24-well plate format, using 0.5 µg plasmid/well and the recommended protocols for each reagent. GFP expression was analyzed 48 hr posttransfection. Each condition was tested in triplicate, and the data points show the mean transfection efficiency + SD.

Improve genome editing outcomes

Lipofectamine® 3000 reagent was developed to break through the boundaries of delivery and facilitate new technologies, such as genome engineering, in more biologically relevant systems. With this new reagent, GeneArt® TALs and CRISPR target the AAVS1 locus in HepG2 and U2OS cells, and show improved transfection efficiency, mean fluorescence intensity, and genomic cleavage. These advancements in delivery help minimize painstaking downstream workflows, enable easier stem cell manipulation, and enhance site-specific insertion of transgenes into the cellular genome.

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Figure 5. Cleavage efficiency of GeneArt® TALs and CRISPRs. The TALs and CRISPRs targeted the AAVS1 locus in (A) U2OS and (B) HepG2 cell lines. Cleavage was assayed using a GeneArt® Genomic Cleavage Detection Kit. 



Figure 6. Transfection efficiency and protein expression using a CRISPR vector. The vector contained an OFP reporter gene and was transfected with Lipofectamine® 2000 or Lipofectamine® 3000 reagent into (A) U2OS and (B) HepG2 cell lines. Bar graphs show reporter gene expression; images show fluorescence of corresponding cells expressing OFP.

*No purchase necessary. This promotion is available only to life science professionals in the US (excluding Puerto Rico) and Canada who are 21 years or older. Sample requests must be received by June 30, 2014, or until sample supplies are depleted, whichever comes first. The sample size is 0.1 mL. Limit one free sample per customer. Health care professionals may not participate in this promotion. Cannot be combined with other discounts or promotions. Offer void where prohibited, licensed, or restricted by federal, state, provincial, or local laws or regulation or agency/institutional policy. Other restrictions may apply.