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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:
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).
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.
Figure 2. The Lipofectamine 3000 transfection protocol was developed to be easy to use, while still ensuring optimum performance and reliability in a wide panel of cell lines.
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.
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.
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.
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%
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.
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.
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 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.
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.
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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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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