SureFECT™ Transfection Reagent

The SureFECT Transfection Reagent is the high-efficiency, low-toxicity solution for the transfection of a wide variety of mammalian cell types. SureFECT is specifically optimized for reverse transfection in normal serum-containing medium, not serum-free medium like many other commercial transfection reagents. Reverse transfection saves an entire day over traditional transfection by plating cells directly into wells and medium already containing reagent-DNA complexes. Utilizing reverse transfection has been reported to produce equivalent or improved transfection efficiencies over traditional methods*. We have also observed better well-to-well reproducibility with reverse transfection protocols. The simplicity of reverse transfection with the SureFECT Transfection Reagent allows you to quickly analyze gene function in your cell line of interest.

Product Listing

Product Name	Cat #	Unit
SureFECT™ Transfection Reagent	SA-01	0.5 ml (enough for 1400 transfections or 12 X 96 well plates)

Why the SureFECT Transfection Reagent?

• Optimized for Reverse Transfection:
  Shortens experiment timeline by a day - plate and transfect cells at the same time.
• High Efficiency:
  Reverse transfect a high percentage of cells.
• Low Toxicity:
  SureFECT reagent is very gentle on cells.

HOW IT WORKS

Performance and Applications Data

Performance Data

Figure 1: SureFECT Reverse Transfects with Greater Efficiency than All Other Traditional Transfection Reagents. In 96-well plates, 50 µL of diluted DNA (0.33 µg pCMVb-Gal plasmid) was mixed with 50 µL dilutions of eight different commercial transfection reagents in serum-free Opti-MEM. COS7 cells (15,000 in 50 µL) in normal medium containing 5% FBS were then added to the wells for reverse transfection. Media was changed 24 h post-transfection. Beta-gal enzymatic activity (OD₅₇₀) was assayed 32 h post-transfection utilizing 0.5 mg/ml CRPG as substrate.

Figure 2: SureFECT Maintains the Viability of Efficiently Reverse Transfected Cells. In 96-well plates, 50 µL of diluted DNA (0.33 µg pCMVb-Gal plasmid) was mixed with 50 µL dilutions containing four different amounts of eight different commercial transfection reagents in serum-free Opti-MEM. COS7 cells (15,000 in 50 µL) in normal medium containing 5% FBS were then added to the wells for reverse transfection. Media were changed 24 h post-transfection. Viability was measured 32 h post-transfection utilizing an acidic phosphatase assay.

Figure 3: SureFECT Works Equally Well On Multiple Cell Lines Commonly Used For Gene Function Studies. SureFECT (0.3 µL per well) was used to reverse transfect MAPK1 siRNA (2 pmole) into different cell types in a 96-well plate. MAPK1 mRNA levels were measured 48 h after transfection using quantitative real-time RT-PCR. The knockdown efficiency (versus a negative control siRNA) is calculated via the ΔΔC_t method.

Application Examples

The reagent performs well in the reverse transfection of nucleic acids such as the SureSilencing siRNA Arrays, the Cignal™ Reporter Assays, the SureSilencing shRNA Plasmids, and other plasmids or constructs.

SureSilencing siRNA Arrays

SureFECT is an exceptional reverse transfection reagent for carrying out siRNA Array cell-based assays
HEK293 H cells expressing the Cignal NFκB Reporter were reverse transfected in the Human NFκB Signaling Pathway siRNA Array. 48 h post-transfection, cells were treated with 50 ng/ml TNFα for 5 h, relative NFκB activity was analyzed by luminescence (red bars), and gene knockdown was also determined by qRT-PCR (blue symbols).

Cignal Pathway Activity Reporter Assays and shRNA Studies

SureFECT is an exceptional reverse transfection reagent for evaluating RNA interference effects with Cignal Reporter Assays
SureFECT was used to transfect 293 H cells with NFκB Cignal Reporter, negative control, and positive control, along with NFκB1siRNA, RelA shRNA, negative control siRNA or negative control shRNA. After 16 hours of transfection, medium was changed to complete growth medium. Dual-Luciferase assay was performed 72 hours post-transfection, and relative firefly luciferase values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were done in triplicates, and the standard deviation is indicated.

* Ziauddin J, Sabatini DM (2001) Microarrays of cells expressing defined cDNAs. Nature 411:107-110.