Lipofectamine 3000 Transfection Reagent product photo

Lipofectamine 3000 Transfection Reagent delivers a high efficiency for hard-to-transfect cell lines. You can achieve over 70% transfection efficiency in a cost-effective way. Explore the benefits of Lipofectamine 3000 reagent:

  • Superior transfection efficiency—10-fold high efficiency into the broadest spectrum of difficult-to-transfect cells
  • Gentle with low toxicity—for improved cell viability
  • Great value—just pennies per reaction for best-in-class transfection results

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Superior transfection performance across a range of hard-to-transfect cell line

When performing transfections in hard-to-transfect cell lines, Lipofectamine 3000 reagent can be an ideal choice. Lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology to enable exceptional transfection performance and reproducible results. It delivers exceptional transfection efficiency with improved cell viability for the widest range of hard-to-transfect and common cell types including HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines (Figure 1).

Fluorescent micrographs showing GFP transfection in cells transfected using Lipofectamine 3000 reagent, Lipofectamine 2000 reagent, and FuGene HD. Cells transfected with Lipofectamine 3000 reagent show the greatest GFP activity.

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 post-transfection. Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines.


A gentle reagent with low transfection toxicity

All four steps in the transfection process were optimized and our most advanced lipid nanoparticle technologies were combined. This superior transfection performance allows you to reduce the reagent dose, thus helping lower toxicity risk when transfecting your cell line.

Graph comparing transfection efficiencies at increasing doses for each transfection reagent, Lipofectamine 2000, Lipofectamine 3000, and Lipofectamine LTX.
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 post transfection 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.

A gentle reagent for CHO cell transfection

Chinese hamster ovary (CHO) is one of the most widely used non-human mammalian cell lines. They are easy to culture and are commonly used in transfection and large-scale production of therapeutic proteins. Lipofectamine 3000 reagent is a specially formulated gentle transfection reagent that is optimized for plasmid delivery into CHO cells with high efficiency and low cell toxicity. The reagent allows for transfection to be performed both in presence or absence of serum. The easy protocol, exceptional plasmid DNA delivery, and the ability to effectively generate stable cell lines makes Lipofectamine 3000 reagent, the perfect reagent for CHO cell transfection experiments.


Increase transfection efficiency and reproducibility with Lipofectamine 3000 reagent


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Comparison between Lipofectamine 3000 and Lipofectamine 2000 reagents

Using the most biologically relevant cell lines helps provide the most relevant answers to your research questions. The exceptional cell spectrum and high transfection efficiency of Lipofectamine 3000 reagent enables excellent flexibility to use your cell line of choice in your studies.

Compare Lipofectamine 3000 reagent and Lipofectamine 2000 reagent in a variety of cell lines using the comparison charts below. Find Lipofectamine 3000 reagent transfection efficiency in each cell line as well as fold protein expression improvement when using Lipofectamine 3000 over Lipofectamine 2000 reagent.

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

Cell typeLipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
NIH/3T34Mouse embryonic fibroblast cell line
4T12Mouse breast tumor cell line
A4312Human epidermoid carcinoma cell line
A5493Human lung cancer cell line
ACHN2Human renal cancer cell line
bEnd.39Mouse brain endothelioma cell line
BJ3Human skin fibroblast cell line
BT-5494Human breast cancer cell line
C2C123Mouse myoblast cell line
C65Rat glioma cell line
Caco-22Human colon carcinoma cell line
Caki-14Human kidney clear cell carcinoma cell line
CHO-K11Chinese hamster ovary cell line
CHO-S1Chinese hamster ovary cell line
COLO 2054Human colon cancer cell line
COS-74Monkey fibroblast-like cell line

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

Cell typeLipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
DU 1452Human prostate cancer cell line
H4603Human lung cancer cell line
H9c23Rat cardiomyocyte cell line
HCC19375Human breast cancer cell line
HCT 1161Human colon carcinoma cell line
HEK-2932Human embryonic kidney cell line
HeLa3Human cervical cancer cell line
Hep 3B2Human liver cancer cell line
Hepa 1-61Mouse liver cancer cell line
Hep G29Human liver cancer cell line
Hs 578T3Human breast cancer cell line
HT-10801Human fibrosarcoma cell line
HT-291Human colon cancer cell line
Huh-74Human hepatoma cell line
Jurkat1Human T lymphocyte cell line

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

Cell type (cont.)
Lipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
K-5621Human myelogenous leukemia cell line
L68Rat skeletal muscle cell line
L9292Mouse fibrosarcoma cell line
LNCaP6Human prostate cancer cell line
MCF 10A5Human mammary epithelial cell line
MCF72Human breast cancer cell line
MDA-MB-2313Human breast cancer cell line
MDA-MB-4531Human breast cancer cell line
MDA-MB-4689Human breast cancer cell line
MDCK1Canine kidney cell line
Neuro-2a1Mouse neuroblastoma cell line
NCI-H232Human lung adenocarcinoma cell line
NCI-H46017Human lung cancer cell line
P191Mouse embryonal carcinoma cell line
PANC-13Human epithelioid carcinoma cell line
PC-122Rat pheochromocytoma cell line

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

Cell type (cont.)
Lipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
RAW 264.74Mouse macrophage cell line
RBL2Rat basophil leukemia cells
RD4Human rhabdomyosarcoma cell line
Saos-24Human osteosarcoma cell line
SH-SY5Y1Human neuroblastoma cell line
SK-BR-34Human breast cancer cell line
SK-MEL-282Human melanoma cell line
SK-N-SH6Human neuroblastoma cell line
SK-OV-33Human ovarian carcinoma cell line
SW4802Human colon cancer cell line
SW6205Human colon cancer cell line
T98G4Human glioblastoma cell line
U2OS3Human osteosarcoma cell line
U-9372Human histiocytic leukemia cell line
Vero1Monkey kidney cell line

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

Cell typeLipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
NIH/3T34Mouse embryonic fibroblast cell line
4T12Mouse breast tumor cell line
A4312Human epidermoid carcinoma cell line
A5493Human lung cancer cell line
ACHN2Human renal cancer cell line
bEnd.39Mouse brain endothelioma cell line
BJ3Human skin fibroblast cell line
BT-5494Human breast cancer cell line
C2C123Mouse myoblast cell line
C65Rat glioma cell line
Caco-22Human colon carcinoma cell line
Caki-14Human kidney clear cell carcinoma cell line
CHO-K11Chinese hamster ovary cell line
CHO-S1Chinese hamster ovary cell line
COLO 2054Human colon cancer cell line
COS-74Monkey fibroblast-like cell line

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

Cell typeLipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
DU 1452Human prostate cancer cell line
H4603Human lung cancer cell line
H9c23Rat cardiomyocyte cell line
HCC19375Human breast cancer cell line
HCT 1161Human colon carcinoma cell line
HEK-2932Human embryonic kidney cell line
HeLa3Human cervical cancer cell line
Hep 3B2Human liver cancer cell line
Hepa 1-61Mouse liver cancer cell line
Hep G29Human liver cancer cell line
Hs 578T3Human breast cancer cell line
HT-10801Human fibrosarcoma cell line
HT-291Human colon cancer cell line
Huh-74Human hepatoma cell line
Jurkat1Human T lymphocyte cell line

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

Cell type (cont.)
Lipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
K-5621Human myelogenous leukemia cell line
L68Rat skeletal muscle cell line
L9292Mouse fibrosarcoma cell line
LNCaP6Human prostate cancer cell line
MCF 10A5Human mammary epithelial cell line
MCF72Human breast cancer cell line
MDA-MB-2313Human breast cancer cell line
MDA-MB-4531Human breast cancer cell line
MDA-MB-4689Human breast cancer cell line
MDCK1Canine kidney cell line
Neuro-2a1Mouse neuroblastoma cell line
NCI-H232Human lung adenocarcinoma cell line
NCI-H46017Human lung cancer cell line
P191Mouse embryonal carcinoma cell line
PANC-13Human epithelioid carcinoma cell line
PC-122Rat pheochromocytoma cell line

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

Cell type (cont.)
Lipofectamine 3000 reagent transfection efficiencyFold protein expression improvement, Lipofectamine 3000 vs 2000 reagentCell type origin
RAW 264.74Mouse macrophage cell line
RBL2Rat basophil leukemia cells
RD4Human rhabdomyosarcoma cell line
Saos-24Human osteosarcoma cell line
SH-SY5Y1Human neuroblastoma cell line
SK-BR-34Human breast cancer cell line
SK-MEL-282Human melanoma cell line
SK-N-SH6Human neuroblastoma cell line
SK-OV-33Human ovarian carcinoma cell line
SW4802Human colon cancer cell line
SW6205Human colon cancer cell line
T98G4Human glioblastoma cell line
U2OS3Human osteosarcoma cell line
U-9372Human histiocytic leukemia cell line
Vero1Monkey kidney cell line

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


Enhance your cancer research

For exceptional performance, utilize Lipofectamine 3000 to enhance transfection in your cancer research experiments. This advanced and versatile reagent works well for transfection in breast, lung, prostate, and liver cancer cells. There are step-by-step protocols to help ensure high transfection efficiency with low toxicity when a specific cancer cell line is coupled with Lipofectamine 3000 transfection reagent.

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.

Graph demonstrating improved transfection efficiencies for Lipofectamine 3000 compared to Lipofectamine 2000 across 17 distinct cell lines.
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 post-transfection. Each condition was tested in triplicate, and the data points show the mean transfection efficiency + SD.


Transfect stem cells or generate induced pluripotent stem cells (iPSCs) with Lipofectamine 3000 reagent

The exceptional 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. 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.

Panel of fluorescent and brightfield micrographs demonstrating transfection efficiency of GFP in H9 ESCs(image A) and iPSCs (image B).

Figure 6. 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.


Lipofectamine 3000 reagent helps improve genome editing outcomes

Lipofectamine 3000 reagent helps break through the boundaries of delivery and facilitates new technologies, such as genome engineering, in more biologically relevant systems. With this reagent, TALENs 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.

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Figure 7. 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 Invitrogen GeneArt Genomic Cleavage Detection Kit.

Two panels of gel images and corresponding bar graphs demonstrating cleavage and gene editing efficiency when comparing Lipofectamine 3000 to Lipofectamine 2000 for delivery of gene editing tools like TALEN and CRISPR sequences.

Figure 8. 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.

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Resources

Our Lipofectamine reagent–specific protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. Search this library to find protocols and citations curated to fit your experimental needs. Filter by cell type, payload, and product of choice to easily unlock your answers.

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