PolyFect Transfection Reagent

Protocol tips

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	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Luciferase assay
The day before transfection, CHO and HEK293 cells were plated in a 96-well dish at 2 × 104 or 6 × 104 per well, respectively. Transient transfection was performed using the Polyfect transfection reagent (Qiagen, Chatsworth, CA). Cells were transfected with 0.25–0.5 μg of endothelial leukocyte molecule-1 (ELAM-1) firefly luciferase construct, 0.025–0.05 μg of the thymidine kinase Renilla luciferase (pRL-TK) vector (Promega, Madison, WI), and 0.25–0.5 μg of the expression vector(s) per well. CHO cells were washed in PBS 2–3 h after transfection. Twenty hours after transfection, cells were stimulated for 4 h. The cells were lysed in passive lysis buffer and the luciferase activity was determined using the dual-luciferase reporter assay system (Promega).

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Manufacturer protocol

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