Lipofectamine 3000 Reagent

WARNING: you may be paying more for less transfection efficiency

Achieve over 70% transfection efficiency in difficult cells for just cents per reaction.

  • Superior efficiency—10-fold higher efficiency into the broadest spectrum of difficult-to-transfect cells
  • Gentle with low toxicity—for improved cell viability
  • Best value—only $0.08* per reaction for best-in-class transfection results

*in USD, based on a 96-well plate comparison

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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.

Lipofectamine 3000 reagent performance

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.

 Download the Lipofectamine 3000 reagent protocol

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Gentle with low toxicity

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.

Lipofectamine 3000 reagent efficiency


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.

Find the cell-specific protocol

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 lines #–C
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 lines D–J
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 lines K–P
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 lines R–Z
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%

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.

Transfection efficiency in cancer cells

 


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.

Reprogramming efficiency of Lipofectamine 3000

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.

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lipo-3000_figure-5

Figure 6. 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.

lipo-3000_figure-6

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

Transfection of 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.

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Packaging must protect the product inside—but that doesn’t have to mean sacrificing sustainability. Lipofectamine products are shipped at ambient temperature, diverting 24,000 ft3 of Expanded Polystyrene (EPS) coolers from landfills annually. We reduce the carbon footprint from packaging and shipping these products by 80 tons (CO2 equivalents).

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