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

See application notes for Invitrogen Lipofectamine 3000 Transfection Reagent

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Lipofectamine 3000 Reagent

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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.

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

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.


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.

Cell type Lipofectamine 3000 reagent transfection efficiency Fold protein expression improvement, Lipofectamine 3000 vs 2000 reagent Cell type origin
NIH/3T3 4 Mouse embryonic fibroblast cell line
4T1 2 Mouse breast tumor cell line
A431 2 Human epidermoid carcinoma cell line
A549 3 Human lung cancer cell line
ACHN 2 Human renal cancer cell line
bEnd.3 9 Mouse brain endothelioma cell line
BJ 3 Human skin fibroblast cell line
BT-549 4 Human breast cancer cell line
C2C12 3 Mouse myoblast cell line
C6 5 Rat glioma cell line
Caco-2 2 Human colon carcinoma cell line
Caki-1 4 Human kidney clear cell carcinoma cell line
CHO-K1 1 Chinese hamster ovary cell line
CHO-S 1 Chinese hamster ovary cell line
COLO 205 4 Human colon cancer cell line
COS-7 4 Monkey fibroblast-like cell line

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

Cell type Lipofectamine 3000 reagent transfection efficiency Fold protein expression improvement, Lipofectamine 3000 vs 2000 reagent Cell type origin
DU 145 2 Human prostate cancer cell line
H460 3 Human lung cancer cell line
H9c2 3 Rat cardiomyocyte cell line
HCC1937 5 Human breast cancer cell line
HCT 116 1 Human colon carcinoma cell line
HEK-293 2 Human embryonic kidney cell line
HeLa 3 Human cervical cancer cell line
Hep 3B 2 Human liver cancer cell line
Hepa 1-6 1 Mouse liver cancer cell line
Hep G2 9 Human liver cancer cell line
Hs 578T 3 Human breast cancer cell line
HT-1080 1 Human fibrosarcoma cell line
HT-29 1 Human colon cancer cell line
Huh-7 4 Human hepatoma cell line
Jurkat 1 Human T lymphocyte cell line

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

Cell type (cont.)
 Lipofectamine 3000 reagent transfection efficiency Fold protein expression improvement, Lipofectamine 3000 vs 2000 reagent Cell type origin
K-562 1 Human myelogenous leukemia cell line
L6 8 Rat skeletal muscle cell line
L929 2 Mouse fibrosarcoma cell line
LNCaP 6 Human prostate cancer cell line
MCF 10A 5 Human mammary epithelial cell line
MCF7 2 Human breast cancer cell line
MDA-MB-231 3 Human breast cancer cell line
MDA-MB-453 1 Human breast cancer cell line
MDA-MB-468 9 Human breast cancer cell line
MDCK 1 Canine kidney cell line
Neuro-2a 1 Mouse neuroblastoma cell line
NCI-H23 2 Human lung adenocarcinoma cell line
NCI-H460 17 Human lung cancer cell line
P19 1 Mouse embryonal carcinoma cell line
PANC-1 3 Human epithelioid carcinoma cell line
PC-12 2 Rat pheochromocytoma cell line

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

Cell type (cont.)
 Lipofectamine 3000 reagent transfection efficiency Fold protein expression improvement, Lipofectamine 3000 vs 2000 reagent Cell type origin
RAW 264.7 4 Mouse macrophage cell line
RBL 2 Rat basophil leukemia cells
RD 4 Human rhabdomyosarcoma cell line
Saos-2 4 Human osteosarcoma cell line
SH-SY5Y 1 Human neuroblastoma cell line
SK-BR-3 4 Human breast cancer cell line
SK-MEL-28 2 Human melanoma cell line
SK-N-SH 6 Human neuroblastoma cell line
SK-OV-3 3 Human ovarian carcinoma cell line
SW480 2 Human colon cancer cell line
SW620 5 Human colon cancer cell line
T98G 4 Human glioblastoma cell line
U2OS 3 Human osteosarcoma cell line
U-937 2 Human histiocytic leukemia cell line
Vero 1 Monkey 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.

 


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.

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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.

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.

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