SiFAT4 (SR312478) and SR312478-FAT4 overexpression plasmid were obtained from Origene, siRNA (A01001) was purchased from Gene pharma. LS174T and SW-620 cell lines were cultured in 6-well plates (5×105 cells per well) to reach confluence of 60% to 80%. Lipofectamine TM 2000 transfection reagent (#11668019, ThermoFisher, USA) was used according to its instruction. A total volume of 75 µg siFAT4 or siRNA was prepared and added to LS174T cells, while FAT4 and an empty plasmid was respectively added to SW-620 cells, and the cell lines were all cultured in DMEM medium. After culturing the cells in CO2 for 6 h at 37 °C, DMEM medium was replaced by 10% FBS/DMEM medium to continue the culture for another 24 h.
Lipofectamine tm 2000 transfection
Manufactured by Thermo Fisher Scientific
Sourced in United States
Lipofectamine TM 2000 is a cationic lipid-based transfection reagent used for the delivery of genetic material, such as DNA or RNA, into eukaryotic cells. It facilitates the uptake and expression of the transfected material within the target cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using lipofectamine tm 2000 transfection
1
Modulating FAT4 Expression in Colorectal Cancer
2
Transient Transfection of HEK293 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293 cells (provided by Dr. Makoto Tominaga (National Institutes of Natural Sciences, Japan)) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum at 37 °C under a humidified atmosphere including 5% CO2. For cell surface staining and Ca2+ imaging, cells were seeded onto a 35-mm chamber (ibidi, Martinsried, Germany). After incubation for 24 h, the TAS1Rs and Gα16-gust44 (0.9 μg of each) were transiently cotransfected into HEK293 cells using LipofectamineTM 2000 transfection reagent (Thermo Fisher Scientific, Waltham, MA, USA; 2.5 μL per 0.9 μg DNA).
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
Revolutionizing how scientists
search and build protocols!