- Versatile — works with a broad range of cell lines
- Reproducible — produces consistent results, lot-to-lot, plate-to-plate, and well-to-well
- Efficient — performs with high siRNA transfection efficiency, allowing use of low siRNA concentrations to minimize nonspecific effects
- Fast — reverse-transfects cells as they are plated, saving a day
A Better Transfection Agent: siPORT™ NeoFX™
Gene silencing experiments often call for critical, but time-consuming optimization experiments. siPORT NeoFX Transfection Agent refines siRNA transfection protocols resulting in less optimization. siPORT NeoFX’s lipid-based formulation can be used to efficiently transfect adherent cells as they are subcultured — without increased cytotoxicity. From start to finish, successful gene silencing experiments can be completed in as early as 24 hours. This streamlined protocol can be adapted to a wide range of cells and experimental designs, including high-throughput applications.
Reverse-Transfect Cells as They are Plated, Saving a Day
Just add siPORT NeoFX to your diluted siRNA, incubate to form transfection complexes, add the complexes to the culture wells, and overlay with cells (Figure 2). This new streamlined protocol, named "neofection", will save researchers a full day of valuable time when compared to traditional plated transfection procedures. The transfection complexes are active and stable even in the presence of serum, so there is no need to remove or replace media following transfection.
Optimized for Lower siRNA Concentrations
Recent reports indicate that siRNA concentrations of 100 nM or higher can lead to nonspecific changes in gene expression (off-target effects) in mammalian cultured cells. Reducing the amount of siRNA used for transfections to 1-20 nM minimizes these nonspecific effects, while still providing extensive silencing of the target gene. siPORT NeoFX Transfection Agent efficiently transfects low concentrations of siRNA. For example, high levels of siPORT NeoFX-mediated gene silencing (>80%) were achieved by transfection of as little as 0.1-10 nM siRNA (targeting GAPDH) into HeLa cells (Figure 3).
Plate-to-Plate, Well-to-Well, and Lot-to-Lot Consistency in Performance
A good transfection agent is expected to reliably deliver high performance among multiple wells for replicate samples and between different lots of the transfection agent. Such characteristics are especially important in high throughput screening of siRNA libraries arrayed in 96 well plates. As shown in Figure 4, there was negligible variability in gene silencing efficiency between multiple wells and multiple plates when siPORT NeoFX was used. Further, three different lots of siPORT NeoFX were found to give very consistent knockdown efficiency (data not shown).
Compatible with a Broad Range of Cell Lines
siPORT NeoFX has been used successfully to deliver siRNA into many cell types, including MCF-7 (human breast adenocarcinoma), HT-29 (human colorectal adenocarcinoma), HeLa-S3 (human cervix adenocarcinoma), HeLa (human cervix adenocarcinoma), HEP-G2 (human hepatocellular carcinoma), BJ (human foreskin fibroblast), A549 (human lung carcinoma), UMR106 (rat osteosarcoma), SKOv3 (human ovarian carcinoma), and SKNAS (human bone marrow neuroblastoma).
siPORT NeoFX Transfection Agent is available in sizes of 0.4 ml and 1.0 ml, which is enough to perform up to 800 and 2000 transfections, respectively, in 96-well tissue culture plates. Instructions for transfection optimization are provided in the detailed Protocol accompanying the transfection agent.
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