Short interfering RNA (siRNA) can be directly introduced into cells by transfection. A lipid-based transfection reagent, such as X-tremeGENE siRNA Transfection reagent, can provide a convenient, reliable, and efficient vehicle for delivering siRNAs into animal cells, enabling the study of cellular and functional consequences of gene knockdown.
This innovative reagent forms a complex with siRNA, as well as with mixtures of siRNA and plasmid DNA (cotransfection), and efficiently delivers the nucleic acids into animal cells to induce gene silencing. Transfection is achieved in just a few steps: mix and incubate diluted transfection reagent with diluted siRNA, then add this complex to cells. Because X-tremeGENE siRNA Transfection Reagent functions exceptionally well in the presence or absence of serum and demonstrates low cytotoxicity, it can be used without media changes. The product is animal-component free.
Solution filtered through 0.2 μm pore size membrane, supplied in polypropylene tubes.
X-tremeGENE siRNA Transfection Reagent efficiently delivers short interfering RNA (siRNA) into many commonly used cell types including HeLa, NIH 3T3, HEK-293, CHO-K1, and COS-7, and several hard-to-transfect cell lines, such as HT29, a human adenocarcinoma cell line.
Caractéristiques et avantages
- Knock down gene expression over 90% in many different cell types.
- Maximize experimental flexibility with a single reagent for siRNA- and cotransfection-based gene-knockdown experiments.
- Produce meaningful results using a reagent that exhibits low cytotoxicity, ensuring that the cellular effects you observe are due to the transfected siRNA rather than the transfection procedure.
- Work with or without serum, avoiding medium changes (e.g., to serum-free medium) before or after transfection.;
X-tremeGENE siRNA Transfection Reagent is a proprietary blend of lipids and other components, free of animal products.
Activity assay: X-tremeGENE siRNA Transfection Reagent (1 - 2.5 μl) is combined with siRNA (0.1 - 0.35 μg) that is specific for the HPRT housekeeping gene. The mixture is used to transfect HEK-293 cells (in a monolayer, 30 - 50% confluent) in the presence of 10% fetal bovine serum (FBS). Following transfection, the decrease of HPRT mRNA cells is determined with the LightCycler® Real-Time PCR System. A knockdown efficiency of 70 - 95% is typically observed when mRNA is measured.
Cytotoxicity analysis: HEK-293 cells are exposed to siRNA/X-tremeGENE siRNA Reagent complexes for 72 hours in the presence of serum, without a media change. Cytotoxicity is then tested by analyzing the cells with the WST-1 Cell Proliferation Reagent (Roche).
For life science research only. Not for use in diagnostic procedures.
X-tremeGENE is a trademark of Roche