Log In Sign up
The largest database of trusted experimental protocols

Lipofectamine 2000

Manufactured by OriGene
Visit Supplier
Sourced in United States

Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for the delivery of nucleic acids, such as plasmid DNA and siRNA, into mammalian cells. It facilitates the uptake of these molecules into the cells, enabling the study of gene expression and function.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (12 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions) Ps100001, by OriGene (2 mentions) Klf4 plasmid, by OriGene (2 mentions) Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions) Oligofectamine, by Thermo Fisher Scientific (1 mentions) C myc, by New England Biolabs (1 mentions) Dual glo luciferase, by Promega (1 mentions) Q5 site directed mutagenesis kit, by New England Biolabs (1 mentions) Luciferase assay kit, by Promega (1 mentions) Recombinant human tnf α, by Thermo Fisher Scientific (1 mentions) Hmgcs2, by Thermo Fisher Scientific (1 mentions) On targetplus smartpool sirna, by Horizon Discovery (1 mentions) Hydrogen peroxide h2o2, by Merck Group (1 mentions) Dld 1, by American Type Culture Collection (1 mentions) Caco 2, by American Type Culture Collection (1 mentions) Shrna targeting, by OriGene (1 mentions) Mir 18a mimic, by RiboBio (1 mentions) Mir 18a mimic, by OriGene (1 mentions) Mir 21 mimic, by RiboBio (1 mentions)

27 protocols using lipofectamine 2000

1

MicroRNA Regulation of Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
For transient transfection of miRNAs, cells at 50% confluence were transfected with either 100 nM pre-miR-24, scrambled pre-miR-24, or anti-miR-24 (Ambion, Life Technologies) in Oligofectamine (Thermo Fisher Scientific). To overexpress BimL, cells were transfected with 3 μg BimLcDNA, a kind gift of Dr. Douglas Green (St. Jude Children's Research Hospital, Memphis, TN, USA), in Lipofectamine 2000 (Thermo Fisher Scientific). To overexpress FIH-1, cells were transfected with 3 μg FIH1 cDNA (OriGene Technologies, Rockville, USA) in Lipofectamine 2000.
Roscigno G., Puoti I., Giordano I., Donnarumma E., Russo V., Affinito A., Adamo A., Quintavalle C., Todaro M., Vivanco M.D, & Condorelli G. (2017). MiR-24 induces chemotherapy resistance and hypoxic advantage in breast cancer. Oncotarget, 8(12), 19507-19521.
+ Open protocol
+ Expand
2

c-MYC Transcriptional Reporter Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
HEK293T cells were seeded at 35,000 cells/well in a 96-well plate. Cells were transfected with the MYC luciferase reporter according to protocol described above using Lipofectamine 2000 (Invitrogen, 11668019) as a transfection reagent. Cells were also transfected with either FLAG-eGFP, FLAG-tagged wild-type c-MYC (Origene, RC201611) or C171Y cMYC mammalian expression plasmids using Lipofectamine 2000. Cells were transfected with 1 μg/well of corresponding plasmid in a final ratio of 3:1 Lipofectamine 2000:DNA. The c-MYC C171Y mutant plasmid was generated using Q5 site-directed mutagenesis kit according to the manufacturer’s instructions (New England Biolabs, E0554S). Sequencing was confirmed with Quintara Biosciences. 24 hours post-transfection, media was carefully aspirated from each well, and 50 μL of fresh media containing either DMSO or compound was added. After 24 hours of compound treatment, Dual-Glo luciferase (Promega, E2920) detection was performed according to manufacturer’s protocol. Firefly and Renilla luminescence were read on a SpectraMax i3 plate reader. Background luminescence was subtracted using a blank control then Firefly:Renilla was calculated for each individual well. Lysates were also obtained for confirmation FLAG-protein overexpression by Western blotting.
Boike L., Cioffi A.G., Majewski F.C., Co J., Henning N.J., Jones M.D., Liu G., McKenna J.M., Tallarico J.A., Schirle M, & Nomura D.K. (2020). Discovery of a Functional Covalent Ligand Targeting an Intrinsically Disordered Cysteine Within MYC. Cell chemical biology, 28(1), 4-13.e17.
+ Open protocol
+ Expand
3

Examining miR-21 Regulation of PDCD4

Check if the same lab product or an alternative is used in the 5 most similar protocols
Eca109 cells were transfected with two luciferase reporter vectors using Lipofectamine 2000, each of which contains full length of pre-miR-21 sequence and 3′-UTR of PDCD4, respectively (Origene). Luciferase reporter vector with PDCD4 mutant target was transfected in parallel as control. Luciferase activities in the cells were assayed using a luciferase assay kit (Promega).
Liu T., Liu Q., Zheng S., Gao X., Lu M., Yang C., Dai F., Sheyhidin I, & Lu X. (2014). MicroRNA-21 Promotes Cell Growth and Migration by Targeting Programmed Cell Death 4 Gene in Kazakh's Esophageal Squamous Cell Carcinoma. Disease Markers, 2014, 232837.
+ Open protocol
+ Expand
4

Colorectal Cancer Cell Line Manipulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human colorectal cancer epithelial cell lines, HT29, Caco2 and DLD1, were obtained from American Type Culture Collection (ATCC, Manassas, VA). HT29 and Caco2 cells were cultured in McCoy’s 5A medium supplemented with 10% FBS and MEM with 15% FBS, 1% sodium pyruvate and 1% nonessential amino acids, respectively. DLD1 cells were grown in RPMI1640 with 10% FBS. The cells were maintained at 37°C in a humidified 5% CO2 incubator. Transfection with ON-TARGET plus SMARTpool siRNAs (Dharmacon, Lafayette, CO) for non-targeting control (NTC, catalog #, D-001810–10), and HMGCS2 (catalog #, L-010179–00) was performed using Lipofectamine RNAi MAX (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. To establish stable cell lines expressing HMGCS2, pCMV6-Entry-HMGCS2 cDNA clone (RC208128, OriGene Technologies, Rockville, MD) or pCMV6-Entry [empty vector (EV), PS100001, OriGene] were transfected into DLD1 or HT29 cells using Lipofectamine™ 2000 and stable transfectants were obtained following G418 selection. Recombinant human TNFα and rosiglitazone (RGZ; a PPARγ agonist) were purchased from PeproTech (Rocky Hill, NJ) and Tocris Bioscience (Bristol, UK), respectively. βHB, 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate (DCFH-DA) and hydrogen peroxide (H2O2) were obtained from Sigma-Aldrich (St. Louis, MO).
Kim J.T., Napier D.L., Kim J., Li C., Lee E.Y., Weiss H.L., Wang Q, & Evers B.M. (2021). Ketogenesis alleviates TNFα-induced apoptosis and inflammatory responses in intestinal cells. Free radical biology & medicine, 172, 90-100.
+ Open protocol
+ Expand
5

Stable Transfection of NTKL in Hepatocellular Carcinoma

Check if the same lab product or an alternative is used in the 5 most similar protocols
Full-length of NTKL was cloned into PCDNA3.1 and stably transfected into QGY-7703 (NTKL-7703) and MHC-97H cells (NTKL-97H) using lipofectamine2000 (Origene, Rockville, MD). Empty vector-transfected cells (Vec-7703 and Vec-97H) were used as controls. NTKL depleted cells were established by stably transfecting shRNA targeting NTKL (Origene, Rockville, MD) into QGY-7703 cells.
Wang J., Liu M., Chen L., Chan T.H., Jiang L., Yuan Y.F, & Guan X.Y. (2015). Overexpression of N-terminal kinase like gene promotes tumorigenicity of hepatocellular carcinoma by regulating cell cycle progression and cell motility. Oncotarget, 6(3), 1618-1630.
+ Open protocol
+ Expand
6

Transfection Protocol for miR-18a Mimic and KLF4 Plasmid

Check if the same lab product or an alternative is used in the 5 most similar protocols
The miR-18a mimic, inhibitor, and negative control miRNA were purchased from RiboBio (Guangzhou China) and transfected into cells at 100 nM concentrations via Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. KLF4 plasmid (1ug; Origene, Rockville, MD, USA) was also transfected into cells in the presence or absence of the miR-18a mimic by Lipofectamine 2000. After transfection twice in 48 h, cells were used in the subsequent experiments. RT-PCR and Western blotting were used to evaluate the transfection efficacy.
Liu L., Cai X., Liu E., Tian X, & Tian C. (2017). MicroRNA-18a promotes proliferation and metastasis in hepatocellular carcinoma via targeting KLF4. Oncotarget, 8(40), 68263-68269.
+ Open protocol
+ Expand
7

Modulating miR21 and Bcl-2 in Lymphoma

Check if the same lab product or an alternative is used in the 5 most similar protocols
SU-DHL-4 and SU-DHL-8 cells were transfected with miR21 mimics (Riobio, Guangzhou, China) or negative control (Riobio) using lipofectamine 2000 (Invitrogen) following the manufacturer’s instruction. For the knock-down assay, SU-DHL-4, SU-DHL-8 cells, and HUVEC were transfected with Bcl-2 siRNA or control siRNA (Origene, Rockville, MD, USA) using lipofectamine 2000.
Zheng Z., Xu P.P., Wang L., Zhao H.J., Weng X.Q., Zhong H.J., Qu B., Xiong J., Zhao Y., Wang X.F., Janin A, & Zhao W.L. (2017). MiR21 sensitized B-lymphoma cells to ABT-199 via ICOS/ICOSL-mediated interaction of Treg cells with endothelial cells. Journal of Experimental & Clinical Cancer Research : CR, 36, 82.
+ Open protocol
+ Expand
8

Overexpression of cRAF in MIA PaCa-2 cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
MIA PaCa-2 cells were transiently transfected with nonspecific control plasmid (Control cDNA) or with 100 nM cRAF expression plasmid (cRAF cDNA; Cat # SC118323; OriGene, Rockville, MD) for 48 h using Lipofectamine 2000 following the manufacturer’s instructions. Following transfection, cells were replated and treated with CMC2.24 for up to 24 h.
Mallangada N.A., Vargas J.M., Thomas S., DiGiovanni M.G., Vaeth B.M., Nemesure M.D., Wang R., LaComb J.F., Williams J.L., Golub L.M., Johnson F, & Mackenzie G.G. (2018). A novel tricarbonylmethane agent (CMC2.24) reduces human pancreatic tumor growth in mice by targeting Ras. Molecular carcinogenesis, 57(9), 1130-1143.
+ Open protocol
+ Expand
9

Knockdown of HIF-1α in Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fifty nM HIF-1α siRNA (ThermoFisher, 106498) was transferred into cells using Lipofectamine™ RNAiMAX Transfection Reagent (ThermoFisher, 13778150) according to the manufacturer’s instructions. HIF-1α was stably knocked down using short hairpin RNA (shRNA, OriGene, TG320380) using Lipofectamine 2000 according to the manufacturer’s instructions. Cells were then treated with indicated agents for 48 hr after siRNA transfection or shRNA infection (19 , 20 (link)).
Li Z., Zhou W., Zhang Y., Sun W., Yung M.M., Sun J., Li J., Chen C.W., Li Z., Meng Y., Chai J., Zhou Y., Liu S.S., Cheung A.N., Ngan H.Y., Chan D.W., Zheng W, & Zhu W. (2019). ERK regulates HIF-1α-mediated platinum resistance by directly targeting PHD2 in ovarian cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(19), 5947-5960.
+ Open protocol
+ Expand
10

Generating Sox9 Knockdown and Overexpression Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
To establish Sox9 knockdown clones, recombinant lentivirus was generated by co-transfecting 293FT cells with shSox9 from Sigma-Aldrich Mission® shRNA bacterial glycerol stock, Clone IDs: TRCN0000020386 (shSox9#386) and TRCN0000020388 (shSox9#388) or non-target control (NTC) plasmids and packaging plasmid mix (System Biosciences, Mountain View, CA). Viral supernatant was collected and used to infect Huh7 and Hep3B. The infected cells were selected with puromycin (Sigma-Aldrich). For overexpression study, either Sox9 expression construct (SC321884: NM_000346, human cDNA clone) or empty vector (Origene Technologies, Rockville, MD) were transfected into SMMC-7721 using Lipofectamine® 2000.
Leung C.O., Mak W.N., Kai A.K., Chan K.S., Lee T.K., Ng I.O, & Lo R.C. (2016). Sox9 confers stemness properties in hepatocellular carcinoma through Frizzled-7 mediated Wnt/β-catenin signaling. Oncotarget, 7(20), 29371-29386.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!