HEK293 cells were used to develop a stable cell line of PTH1RFRET biosensor. Cells seeded into 100-mm dishes were transfected at a confluence of 60% with 2 µg of cDNA encoding PTH1RFRET with Lipofectamine 3000 Transfection Reagent Kit (Qiagen), according to the manufacturer’s protocol. Transfected clones were selected with 600 μg/mL G-418 (VWR International) and sorted with a flow cytometer. Monoclonal single clones were grown in DMEM supplemented with 200 μg/mL G-418. The best clone was selected for further experiments based on the brightness and amplitude of the saturating PTH stimulation of the PTH1RFRET biosensor in plate reader experiments.
Lipofectamine 3000 transfection reagent kit
Manufactured by Qiagen
Lipofectamine 3000 Transfection Reagent Kit is a laboratory product designed for the efficient transfection of nucleic acids, including DNA and RNA, into a variety of cell types. The kit provides the necessary components to facilitate the delivery of genetic material into cells, enabling researchers to study gene expression, protein function, and other cellular processes.
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Lab products found in correlation
2 protocols using lipofectamine 3000 transfection reagent kit
1
Stable PTH1R FRET Biosensor in HEK293
2
Stable PTH1R FRET Biosensor Cell Line
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HEK293 cells were used to develop a stable cell line of PTH1RFRET biosensor. Cells seeded into 100 mm dishes were transfected at a confluence of 60 % with 2 µg of cDNA encoding PTH1RFRET with Lipofectamine 3000 Transfection Reagent Kit (Qiagen), according to the manufacturer´s protocol. Transfected clones were selected with 600 μg/mL G-418 (VWR International) and sorted with a flow cytometer. Monoclonal single clones were grown in DMEM supplemented with 200 μg/mL G-418. For further experiments, the best clone was selected based on the brightness and amplitude of the saturating PTH stimulation of PTH1RFRET biosensor in plate reader experiments.
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