The ability to successfully transfect a cell varies significantly depending on the cell type, protocol, and the composition of the transfection reagent. X-tremeGENE™ transfection reagents are designed to efficiently transfect common and difficult-to-transfect cells with a variety of molecules, including DNA, small RNAs, and CRISPR/Cas9 components.
Figure 1: X-tremeGENE™ HP Reagent efficiently transfects human primary fibroblasts.
Primary fibroblasts were isolated from human foreskin and transfected with 4 μL X-tremeGENE™ HP Reagent and 1 μg of GFP-encoding plasmid DNA in a 6-well cell culture plate. GFP expression was visualized 48 hours after transfection. The left panel shows GFP fluorescence, and the right panel shows the corresponding brightfield image.
Figure 2: X-tremeGENE™ HP and 9 Reagents outperform the competitor reagent in human mesenchymal stem cells.
Human mesenchymal stem cells were isolated from bone marrow aspirate of four different donors. Cells were transfected using X-tremeGENE™ HP Reagent (XHP), X-tremeGENE™ 9 Reagent (X9), and a competitor reagent (LTX), using the indicated ratios (μL reagent : μg GFP-encoding plasmid DNA. Ratio 2:1 was not tested using X-tremeGENE™ 9 Reagent). Untransfected cells served as control (neg). FACS analysis was performed 48 hours after transfection.
Figure 3.X-tremeGENE™ HP DNA Transfection Reagent outperforms competitor reagents in difficult-to-transfect cell types.
Cells were transfected using X-tremeGENE™ HP Reagent (XHP), and competitor reagents (LTX and L2K), at the indicated ratios (μL reagent : μg GFP-encoding plasmid DNA). Transfection efficiency was visualized using GFP fluorescence microscopy.
Note: The above data was produced using either a GFP-encoding pcDNA3.1 plasmid or a luciferase-encoding pCI plasmid, both with the cytomegalovirus (CMV) promoter. These recommendations are guidelines based on experimental findings. The optimal reagent:DNA ratio must be determined empirically.