Catalog Number Amount of reagent 410-002 0.2 mL 410-015 1.5 mL 410-060 4 x 1.5 mL

1.5 mL of INTERFERin®-HTS is sufficient to transfect 200-300 plates in (96-well) . Bulk sizes available, please contact us.

INTERFERin^®-HTS leads to over 90% cell death in a standard phenotypic cell death assay, using 10 nM of a cell death-inducing PLK1 siRNA (Fig. 1). The assay is significant, as INTERFERin^®-HTS shows minimal cytotoxicity (89% ± 7% cell viability) with non-targeting siRNA.

Figure 1. Highly efficient siRNA-induced cell death in HeLa cells in 384-well plates using INTERFERin®-HTS. HeLa cells were transfected with siRNA inducing cell death (PLK1) and negative control (non-targeting siRNA) following a reverse transfection protocol (10 nM siRNA; 0.05 µL INTERFERin®-HTS per well). Cell viability was measured by automated fluorescent microscopy after DAPI staining 72 hours after transfection (Data kindly provided by the Transfected Cell Arrays Platform IGBMC-CERBM, Illkirch, France).

Transfection of a siRNA targeting endogenous lamin A/C using INTERFERin^®-HTS leads to higher silencing efficiency than the same experiment performed with three other popular transfection reagents (Fig. 2).

Figure 2. siRNA transfection with INTERFERin®-HTS gives higher silencing efficiency than competitor reagents. HeLa cells were transfected in 96-well plates with 10 nM siRNA duplexes targeting lamin A/C following a reverse transfection protocol according to the manufacturer‘s recommendations. Lamin A/C expression was measured 24 hours after transfection by branched DNA assay and normalized to GAPDH expression.

• Easy to use: the reagent is an aqueous solution stored at 4 °C with a shelf-life of one year, compatible with the use of serum and antibiotics.
• Reagent diluted in aqueous solution is stable for more than a week.
• siRNA/INTERFERin^®-HTS complexes are stable up to 4 hours at room temperature (Fig. 3).
• siRNA/reagent complex can be frozen at -80°C at least a week in plates.
• Reverse and forward protocols are provided for 96- and 384-well plates.

Figure 3. siRNA/INTERFERin®-HTS complexes are stable for up to 4 hours at room temperature. siRNA/INTERFERin®-HTS complexes were formed according to the manufacturer‘s recommendations using 10 nM siRNA duplexes and incubated at room temperature for the indicated time. Subsequently, HeLa cells were transfected in 96-well plates with 10 nM siRNA duplexes targeting lamin A/C following a reverse transfection protocol according to the manufacturer‘s recommendations. Lamin A/C expression was measured 24 hours after transfection by branched DNA assay and normalized to GAPDH expression.

INTERFERin^®-HTS gives highly reproducible results at all levels:

• Well to well ► The data presented in Figure 3 were done in triplicate yielding a very small standard deviation and highly significant comparisons (Fig.4).
• One experiment to the next ► Three independent transfection experiments using the same batch of INTERFERin^®-HTS show identical silencing efficiencies (Fig. 4)
• Batch to batch ► Transfection using three different batches of INTERFERin^®-HTS shows highly consistent results (Fig. 5).

With INTERFERin^®-HTS, your results are reliable.

Figure 4. Experiment to experiment reproducibility in 96-well plates. A549 cells stably expressing the luciferase gene were transfected with 10 nM GL3Luc siRNA and negative control duplexes using 0.05 µL of INTERFERin®-HTS in 3 independent experiments. Luciferase expression was measured 48 hours after transfection.

Figure 5. Batch to batch reproducibility in 96-well plates. A549 cells stably expressing the luciferase gene were transfected with 10 nM GL3Luc siRNA duplexes and negative control using 0.05 µL of INTERFERin™-HTS from 3 different batches. Luciferase expression was measured 48 hours after transfection.

Higher inhibition of gene expression is obtained with INTERFERin^®-HTS using lower amounts of reagent than competitors; at least twice as many plates can be transfected with the same vial size (Table 1).

Table 1. Volume of reagent (INTERFERin®-HTS and competitors) required per well in 96-well plates and number of transfected plates with 1.5 mL reagent size, according to the manufacturer’s recommendations for siRNA reverse transfection.

INTERFERin^®-HTS shows superior advantages for siRNA high throughput screening, in particular, very low amounts of reagent are required to perform siRNA transfection. INTERFERin^®-HTS is therefore extremely cost-effective.