PolyFect Transfection Reagent

DNA transfection Mammalian cells - Immortalized cell lines HEK293

Experiment
DNA transfection Mammalian cells - Immortalized cell lines HEK293
Product
PolyFect Transfection Reagent from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
polyFect–DNA complex formation should be performed in the absence of serum, antibiotics, and proteins.

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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Papers

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Paper title
Cutting edge: a Toll-like receptor 2 polymorphism that is associated with lepromatous leprosy is unable to mediate mycobacterial signaling.
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Manufacturer protocol

Download the product protocol from Qiagen for PolyFect Transfection Reagent below.

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