Qiagen
HiPerFect Transfection reagent
301705
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In the present study, pancreatic beta cells from rats were transfected with desired mammalian overexpression plasmid in which overexpressed proteins were tagged with green fluorescent protein at N-terminal.
Plasmid transfection in rat pancreatic beta cells
Rat pancreatic beta cell culture, nuclease free water, overexpression plasmid, HiPerFect reagent, MEM medium, cell culture media with 10% serum, CO2 incubator etc
Rats of 5 weeks age maintained on standard management conditions were sacrificed using standard ethical protocol. Pancreas were removed, pancreatic beta cells were isolated and in vitro cell culture were established. Cultured pancreatic beta cells were transfected with desired overexpression plasmid using HiPerfect reagent obtained from Qiagen. In brief, pancreatic beta cells were passaged 12hr prior to the transfection with a 70% confluence area. For transfection, 1 micg of plasmid (with 8Kb length) were mixed with HiPerFect reagent as per manufacturer instructions, cells were overlayed with above reagent and incubated in a CO2 incubator for 4-5 hrs. After that old media of cultured cells were replaced with fresh media containing 10% serum. Transfected cells were maintained at CO2 incubator for at least 36 hrs to allow overexpression of GFP-labeled proteins.
No
During transfection viability of cells were not affected. Almost 90% of the cells were transfected with overexpression plasmid as quantified under fluorescent microscope for GFP labeled proteins.
Easy to use and highly efficient transfection reagent.
I strongly recommend this product for transfection
Product Name: HiPerFect Transfection Reagent (1 ml)Supplier Page