High transfection efficiency allowing low siRNA concentration
siRNA concentrations above to 100nM can lead to non-specific changes in gene expression (off-target effects) in mammalian cultured cells. Reducing the amount of siRNAs used for transfections down to 1-20 nM minimize these non-specific effects. The powerful ICAFectin™ 442 leads to a high release of siRNA within the cell and allows using low concentration of siRNA, allowing researchers to choose the siRNA concentration they wish to use. It can be used to transfect a wide range of cell types such as HeLa, HEK 293, HepG2, MCF-7, HUVEC, HaCaT cells and many others.
Lamin A/C silencing efficiency using ICAFectin™442 reagent as a function of siRNA concentration. Anti-LaminA/C siRNA amounts ranging from 0.4 to 20nM (3.75 to 200ng/well) were formulated with ICAFectin™442. Results of real-time quantitative RT-PCR analysis of human Lamin A/C mRNA after transfection of HeLa cells showed that ICAFectin™442 reagent is efficient with very low concentration of siRNA.In the data shown above, efficient knockdown (>80 %) is achieved with siRNA concentrations in the range 1-20 nM.
Fluorescence microscopy visualization of GFP silencing and siRNA internalization. The GFP-expressing human lung cancer H1299 cells were transfected with control siRNA (A,B) or 3’-rhodamine labeled anti-GFP siRNA (C-F). The siRNA molecules were formulated in the absence (“naked” siRNA in A,C,E) or in the presence of ICAFectin™442 reagent (B,D,F). The transfected H1299 cells were observed using a FITC filter to visualize GFP fluorescence (A,D) or a rhodamine filter to visualize siRNA internalization (E,F).Easy-to-use procedure
ICAFectin™442 Transfection Reagent is provided as a ready-to-use solution - Simply add the reagent to your diluted siRNA, mix, incubate, and pipet the complexes onto the cells. Transfection can be performed in the presence of serum, eliminating the need to remove complexes from the cells.
Highest efficiency compared to existing transfection products on the market
The knock down efficiency of ICAFectin™ 442 was compared to others products available on the market. ICAFectin™ 442 shows the highest efficiency with a variety of cell lines.
Lamin A/C silencing efficiency using ICAFectin™442 reagent or competitor reagent. Anti-LaminA/C siRNA at 2 and 4nM(18.75 and 37.5 ng/well) were formulated with ICAFectin442 or commercial reagents. Results of real-time quantitative RT-PCR analysis of human Lamin A/C mRNA after transfection of HeLa cells showed that ICAFectin™442 reagent was more efficient than the top leader products on the market.
ICAFectin™ 442 has very low cytotoxicity
Cell viability analysis after transfection using ICAFectin442 or competitor reagents.
Rapid Reverse transfection protocol for high-throughput studies
Transfection with ICAFectin™ 442 Transfection Reagent can be performed using the common transfection procedure of seeding the cells 24 hours before transfection. Alternatively, protocols are provided for reverse transfection in 24-well plates, 96-wells plate and 384-well plates (see ICAfectin™ 442 manual). In these protocols, cells are seeded and transfected on the same day. Reverse transfection protocols are rapid and convenient, can easily be automated and are frequently used for high-throughput experiments.