Achieve over 70% transfection efficiency in difficult cells for just cents per reaction.
*in USD, based on a 96-well plate comparison
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.
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.
| 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%
| 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.
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.
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 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.
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.
For Research Use Only. Not for use in diagnostic procedures.
