Superior efficiency into the broadest spectrum of difficult-to-transfect cell types
Lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology 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 Invitrogen 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 hours posttransfection. Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines.
Figure 2. The transfection protocol for Lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines.
When developing Lipofectamine 3000 reagent, 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 Invitrogen Lipofectamine LTX Reagent.
Enhance your cancer research
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.
Table 1. Lipofectamine 3000 reagent yields higher transfection efficiencies than Lipofectamine 2000 reagent when tested in a variety of cell lines.
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.
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 hours posttransfection. Each condition was tested in triplicate, and the data points show the mean transfection efficiency + SD.
Generate induced pluripotent stem cells (iPSCs)
The superior transfection efficiency of Lipofectamine 3000 reagent in conjunction with Invitrogen Epi5 Episomal iPSC Reprogramming Kit enables highly efficient reprogramming of somatic cells without the need for electroporation.
Figure 5. Reprogramming efficiency of Lipofectamine 3000 reagent compared to electroporation. BJ fibroblasts as well as neonatal (HDFn) and adult (HDFa) human dermal fibroblasts were reprogrammed to iPSCs by transfection of Epi5 vectors using either Lipofectamine 3000 reagent or the Invitrogen Neon Transfection System. Colonies were (A) visualized by brightfield microscopy and (B) stained for alkaline phosphatase.
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 reagent, Invitrogen GeneArt TALs and CRISPR sequences 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.
Figure 7. 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.
Transfect stem cells
Figure 8. Transfection of stem cells. (A) H9 ESCs or (B) iPSCs were transfected using Lipofectamine 3000 reagent. Cells were visualized by fluorescence microscopy and processed using flow cytometry to determine transfection efficiency.
“Excellent!" —Hon-Kit Andus Wong, PhD, Harvard Institutes of Medicine
“It is working very well with good efficiency and low toxicity." —Bassem Khalil, Albert Einstein College of Medicine
“Better than Lipofectamine 2000 and FuGENE!" —Sachin Katyal, PhD, Manitoba Institute of Cell Biology
“Excellent. Far better transfection efficiency than Lipofectamine® 2000 when we compared them head-to-head." —Suzanne Ridges, PhD, University of Kentucky
“We were very happy and surprised to see Lipofectamine® 3000 reagent provide a more than 10-fold difference in transfection efficiency in our difficult-to-transfect cell line. There was even a reduction in cell death. Awesome results! —Rui Eduardo Castro, PhD, University of Lisbon
“Spectacular increase in transfection efficiency and dramatic decrease in cell toxicity. Way ahead of the others!” —Emmanuel Boucrot, PhD, Principal Investigator, University College London
“I tried Lipofectamine® 3000 on several commonly used human and mouse tumor cell lines (293T, HeLa, A549, NIH3T3)….Lipofectamine® 3000 has a reduced cytotoxicity in comparison to Lipofectamine®2000, which is a great improvement to me…” —Dr. Jianming Xu, Researcher at the German Cancer Research Center, Heidelberg, Germany
"Successfully transfected GT1-1 cells. Never achieved with any other reagent except viruses and nucleofectors.” —Julien Sebag, PhD, Assistant Professor, Vanderbilt University
“Absolutely excellent for efficiency and easy to use. It greatly increased the transfection rate in our cell models.” —Vittorio Ricci, MD PhD, University of Pavia
“EXCELLENT!" —Yvonne Hui, University of South Carolina
"Fantastic! Better than any other transfection reagents!" —Lingxiao Li, PhD, University of Arizona
“I found that Lipofectamine® 3000 Reagent kit results in much better transfection efficiency in neonatal cardiomyoctes compared to Lipofectamine® 2000 reagent.” —Researcher, Italy
“Big improvement. High efficiency, low cytotoxicity and more convenient protocol.” —Researcher, Canada
“Much higher efficiency than Lipofectamine® 2000, tested in human H9 ESCs” —Researcher, USA
“Compared to Lipofectamine® 2000, Lipofectamine® 3000 was significantly less cytotoxic! Lipofectamine® 3000 is very excellent!” —Researcher, USA
“It has exceeded my expectations and has worked better than our current reagent (FuGENE® HD) in H9c2 cells.” —Researcher, USA
“Best kit ever.” —Researcher, USA
“Up to now it was the only reagent that worked on transfecting our very difficult cells.” —Researcher, Portugal
“Great product, easy protocol and great efficiency.” —Researcher, Canada
“It greatly increases transfection efficiency from 10% to 80%.” —Researcher, Canada
“It [Lipofectamine® 3000] worked really well on HepG2 cells. Easy to use.” —Researcher, Canada
Lipofectamine 3000 Reagent Video
For Research Use Only. Not for use in diagnostic procedures.