GenJet™ In Vitro DNA Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- For high efficiency, transfect cells at high density. 70~80% confluency is highly recommended - To lower cytotoxicity, transfect cells in presence of serum (10%) and antibiotics

Upstream tips
- For high efficiency, transfect cells at high density. 70~80% confluency is highly recommended - To lower cytotoxicity, transfect cells in presence of serum (10%) and antibiotics

Publication protocol

DNA Transfection and Luciferase Reporter Assay
HLF were plated at 5 × 104 cells/ml and incubated overnight in growth medium. Transient DNA transfections were performed using GenJet reagent (SL100488, SignaGen Laboratories, Gaithersburg, MD) following the standard manufacturer's instructions. Cells were cotransfected with 300 ng of desired firefly luciferase reporter plasmid, 200 ng of constitutively active thymidine kinase promoter-Renilla luciferase reporter plasmid, and either 500 ng of pcDNA3.1 or 500 ng of p3xFLAG-MKL1. Cells were placed in growth media overnight and then serum-starved for 24 h followed by stimulation with the desired agonists for the desired time points as indicated in the figure legends. Cells were then washed with PBS and lysed in protein extraction reagent (78501, Thermo Fisher Scientific). The lysates were assayed for firefly and Renilla luciferase activity using the Dual-luciferase assay kit (E1960, Promega). To account for differences in transfection efficiency, firefly luciferase activity of each sample was normalized to Renilla luciferase activity.

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Manufacturer protocol

Download the product protocol from SignaGen Laboratories for GenJet™ In Vitro DNA Transfection Reagent below.

Download manufacturer protocol

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