siRNA High-throughput Screening

INTERFERin® HTS is a new generation siRNA transfection reagent especially developed for high-throughput screening (HTS) applications providing great silencing efficiency, excellent reproducibility and high cell viability with very low amounts of reagent.

  • Great silencing from 10 to 50 nM siRNA
  • Designed for automated procedures
  • Reproducible at all levels
  • Small volume of reagent per well - cost effective

INTERFERin® HTS is cost-effective, easy to handle, compatible with serum and antibiotics, and comes with reverse and forward protocols for 96- and 384-well plates.

Choose INTERFERin® siRNA tranfection reagent for day-to-day transfections.

Ordering information

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 are available, please contact us to find out more.

Efficient gene silencing with low cytotoxicity using a standard phenotypic assay

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.

INTERFERin HTS

 

Fig. 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).

 

The highest efficiency

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).

 INTERFERin HTS fig4

Fig. 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. 

 


Especially designed for automated procedures

  • 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.

INTERFERin automated

 

Fig. 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. 

 

 




Robust - Reproducible

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.

INTERFERin fig4 (1)Fig. 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.



 

INTERFERin fig 5

Fig. 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. 

 

 


Lower reagent volume per well - more transfections per vial

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).

INTERFERin HTS table

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.