RPTECs were seeded at a density of 1.5 × 105/well in 24-well culture plates. Lipofectamine LTX regent (Invitrogen; Life Technologies) was used for reverse transfection of cells with 500 ng of the pGL4.12 reporter construct or with 200 ng of the pGL4.13 reporter constructs. A total of 10 ng of the pRL-TK vector (Promega) was also transfected as an internal control reporter. Cells were harvested 24 h after transfection and lysates were analyzed using the Dual-Luciferase reporter assay system (Promega). The ratio of firefly (expressed from pGL4.12 reporter constructs) to Renilla (expressed from pRL-TK) luciferase activity in each sample served as a measure of normalized luciferase activity.
Lipofectamine ltx regent
Manufactured by Thermo Fisher Scientific
Lipofectamine LTX is a transfection reagent used for the delivery of nucleic acids, such as plasmid DNA or small interfering RNA (siRNA), into mammalian cells. It is designed to facilitate the uptake and expression of these genetic materials within the target cells.
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Lab products found in correlation
2 protocols using lipofectamine ltx regent
1
Reverse Transfection of Luciferase Reporters
2
Dual-Luciferase Transfection Assay
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RPTECs were seeded at a density of 2.0 × 105/well in 24-well culture plates. Lipofectamine LTX regent (Invitrogen) was used for reverse transfection with 500 ng of the pGL4.25 reporter construct or 200 ng of the pGL4.13 reporter constructs. Ten nanograms of the pRL-TK vector (Promega) was also transfected as an internal control reporter. Cells were harvested 24 h after transfection, and lysates were analyzed using the Dual-Luciferase reporter assay system (Promega) with Berthold Lumat3 LB9508 (Berthold Japan Co, Ltd). The ratio of firefly (derived from pGL4.25 and pGL4.13 reporter constructs) to Renilla (derived from pRL-TK) luciferase activity in each sample served as a measure of normalized luciferase activity.
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