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

Cell culture, fractionation, transfection, microinjection, FISH and immunofluorescence
Cell culture, DNA transfection or microinjection, digitonin extraction, FISH staining and immunostaining were performed as previously described (; ; ). BAT3-knockout MEF cells were obtained from Hitoshi Okada (University of Toronto, Toronto, Canada) (). BAT3 cells were grown in DMEM supplemented with 10% FBS and 2-mercaptoethanol. U2OS and COS7 cells were transfected using GenJet Transfection Reagent (SignaGen Laboratories). BAT3 and MEFs cells were transfected using JetPrime Polyplus (Invitrogen) transfection reagent. Cell fractionation was performed as previously described (). Samples were separated by SDS-PAGE and analyzed by western blotting using rabbit polyclonal antibodies against Trapα (dilution 1:1000; ), Sec61β (dilution 1:1000; ), GAPDH (dilution 1:1000; Abgen) and GFP (dilution 1:1000; Molecular Probes), and monoclonal mouse antibody against α-tubulin (dilution 1:1000; Sigma). To detect t-ftz protein, which contains an HA epitope (), samples were immunoblotted with anti-HA mouse monoclonal antibody (GeneTex; dilution 1:1000).

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Manufacturer protocol

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

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