jetPEI® DNA transfection, HTS application

DNA transfection Mammalian cells - Immortalized cell lines PANC-1

Experiment
DNA transfection Mammalian cells - Immortalized cell lines PANC-1
Product
jetPEI® DNA transfection, HTS application from Polyplus transfections
Manufacturer
Polyplus transfections

Protocol tips

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Transfection and luciferase assay
Transfections were performed using jetPEI (Polyplus-transfection Inc, NY, USA) according to the manufacturer's instructions. 5 x 10HCT116 cells were seeded in 6-well plates for Luc assays. The next day, when the cells were about 50% confluent, co-transfection with 3 μg of PY4-SV40-LUC vector plus 0.3 ng of pRL-CMV (Promega) was performed. 24h after the transfection medium was replaced and the cells were exposed to cetuximab, different concentration of terpinen-4-ol or left untreated. Luc assay was performed 48 h post after the treatment. Briefly, cells were washed once with PBS and then lyzed in 250 ~μl of lysis buffer for 5 min at room temperature. The lysates were centrifuged at 14,000 rpm for 5 min, and 20 μl of each lysate were used to measure Luc reporter gene expression. The Luc activity was normalized to Renilla Luc activity from a parallel co-transfection of pRL-CMVDual Luc system, Promega).

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Papers

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Paper title
Terpinen-4-ol: A Novel and Promising Therapeutic Agent for Human Gastrointestinal Cancers
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Manufacturer protocol

Download the product protocol from Polyplus transfections for jetPEI® DNA transfection, HTS application below.

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