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Lipofectamine 2000 reagent

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Lipofectamine 2000 reagent is a cationic lipid-based transfection reagent used for the delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into mammalian cells. It facilitates the formation of lipid-nucleic acid complexes that can enter cells through endocytosis, enabling efficient gene delivery and expression.

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Lab products found in correlation

Lipofectamine 2000 reagent, by Thermo Fisher Scientific (4 mentions) Hifi dna assembly system, by New England Biolabs (2 mentions) Opti mem medium, by Thermo Fisher Scientific (2 mentions) Luciferase assay system, by Promega (2 mentions) Renilla luciferase assay system, by Promega (2 mentions)

Spelling variants of this product

Lipofectamine 2000 (19 citations) Lipofectamine (3 citations)

2 protocols using lipofectamine 2000 reagent

1

Evaluating UCP1 Variants in hTERT A41hBAT-SVF

Check if the same lab product or an alternative is used in the 5 most similar protocols
The mouse UCP1 (UCP1WT) and the S-nitrosylation resistant (UCP1SNOR) variants were generated in a pcDNA3.1 backbone (Addgene, V790–20) using cDNA isolated from WT BAT. Site-directed mutagenesis was then done with the NEBuilder HiFi DNA assembly system (E5520S) following the manufacturer’s instructions. Primers used for cloning are listed in the Table S1.
The hTERT A41hBAT-SVF were transfected with either pcDNA-EGFP, UCP1WT or UCP1SNOR (2.5 μg/well, pcDNA3.1 backbone) using Lipofectamine 2000 reagent (ThermoFisher Scientific, L3000015) in Opti-MEM medium (ThermoFisher Scientific, 31985–062). At 24–48 hours post-transfection, cells were exposed to isoproterenol (ISO, 1 μM, Sigma, 54750-10-6) in the presence or absence of S-Nitroso-N-acetyl-DL-penicillamine (SNAP, 100 μM, Sigma, N3398) for 2 hours. Cells were then trypsinized, spun at 500xg for 5 minutes at 4°C and assessed for UCP1-dependent oxygen consumption (see below).
HEK293A cells (ATCC, CRL-1573) were transfected with DNA constructs (0.5 μg/well) using Lipofectamine 2000 reagent in Opti-MEM medium. At 48 hours post transfection, the activities of Firefly and Renilla luciferase were measured using the Luciferase Assay System (Promega, E1500) and Renilla Luciferase Assay System (Promega, E2810), separately.
Sebag S.C., Zhang Z., Qian Q., Li M., Zhu Z., Harata M., Li W., Zingman L.V., Liu L., Lira V.A., Potthoff M.J., Bartelt A, & Yang L. (2021). ADH5-mediated NO bioactivity maintains metabolic homeostasis in brown adipose tissue. Cell reports, 37(7), 110003.
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2

Evaluating UCP1 Variants in hTERT A41hBAT-SVF

Check if the same lab product or an alternative is used in the 5 most similar protocols
The mouse UCP1 (UCP1WT) and the S-nitrosylation resistant (UCP1SNOR) variants were generated in a pcDNA3.1 backbone (Addgene, V790–20) using cDNA isolated from WT BAT. Site-directed mutagenesis was then done with the NEBuilder HiFi DNA assembly system (E5520S) following the manufacturer’s instructions. Primers used for cloning are listed in the Table S1.
The hTERT A41hBAT-SVF were transfected with either pcDNA-EGFP, UCP1WT or UCP1SNOR (2.5 μg/well, pcDNA3.1 backbone) using Lipofectamine 2000 reagent (ThermoFisher Scientific, L3000015) in Opti-MEM medium (ThermoFisher Scientific, 31985–062). At 24–48 hours post-transfection, cells were exposed to isoproterenol (ISO, 1 μM, Sigma, 54750-10-6) in the presence or absence of S-Nitroso-N-acetyl-DL-penicillamine (SNAP, 100 μM, Sigma, N3398) for 2 hours. Cells were then trypsinized, spun at 500xg for 5 minutes at 4°C and assessed for UCP1-dependent oxygen consumption (see below).
HEK293A cells (ATCC, CRL-1573) were transfected with DNA constructs (0.5 μg/well) using Lipofectamine 2000 reagent in Opti-MEM medium. At 48 hours post transfection, the activities of Firefly and Renilla luciferase were measured using the Luciferase Assay System (Promega, E1500) and Renilla Luciferase Assay System (Promega, E2810), separately.
Sebag S.C., Zhang Z., Qian Q., Li M., Zhu Z., Harata M., Li W., Zingman L.V., Liu L., Lira V.A., Potthoff M.J., Bartelt A, & Yang L. (2021). ADH5-mediated NO bioactivity maintains metabolic homeostasis in brown adipose tissue. Cell reports, 37(7), 110003.
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