Product Catalog Number Amount of reagent Amount of 150 mM NaCl solution 112-01N 0.1 mL 5 mL FectoFly™ 112-10N 1 mL 50 mL 112-40N 4 x 1 mL 4 x 50 mL

1 ml of FectoFly™ is sufficient to perform 100 to 200 transfections in 6-well plates. For bulk sizes, Please contact us.

By simple and rapid transient transfection method, FectoFly™ provides high transfection efficiency in insect cells and robust protein production over several days (Fig. 1).

Fig 1. Protein production in S2 cells over 5 days using FectoFly transfection reagent. A suspension culture of 106 cells/ml was transfected using 2 µg pCMV-EGFPLuc and 6.4 µl of FectoFly per ml of culture in Insect-XPRESS™ medium (Lonza).

FectoFly™ provides high transfection efficiency in most commonly used insect cells (Fig. 2) allowing efficient protein production.

Figure 2. Comparison of FectoFly™ in various adherent cell lines. Sf9, Sf21, S2 or Tn5 adherent cells were transfected in 24-well plates pCMV-EGFP-Luc plasmid. Luciferase assays were performed 72 h after transfection.

FectoFly™ allows successful transfection of adherent and suspension insect cell cultures both in the latest generation of syntheticserum-free media (Fig.1, 3 – Sf21 and Sf9) and in the presence of serum (Fig.3 – S2). Transfection efficiency ranges routinely from 30% to 50% and cell viability is above 80%.

		Fig 3. GFP expression in Sf21, Sf9 cells grown in suspension, 72 h after transfection of  pCMV-EGFP-Luc plasmid using FectoFly™.

FectoFly™ was compared to other commercially available reagents dedicated to the transfection of insect cells (Fig. 4). As shown for Sf9 cells, the highest protein expression levels were obtained with FectoFly™.

Fig 4. Comparison of FectoFly™ with other insect cell-specific transfection reagents. Sf9 adherent cells were transfected in 6-well plates with pCMV-EGFPLuc plasmid using insect cell transfection reagents according to the supplier’s recommendations.

Transfection of insect cells with FectoFly™ is straightforward. o Fast : 3 steps protocol o Simple: 1 µl of FectoFly™ per µg of DNA, 1:1 ratio o Versatile: efficient in serum-free synthetic media and in the presence of serum Figure 5. FectoFly™ standard protocol.

Catalog Number Amount of reagent 115-010 10 ml 115-375 375 ml

Large sizes are available upon request. Please contact us.

PEIpro™ has been formulated at 1 mg/mL to fit current standard for transfection with PEI. This concentration eases the transfection protocol optimization. PEIpro protocol is available upon request. Contact the Technical Support.

Positively charged reagent/DNA complexes are needed to achieve high transfection efficiency. The global charge of the complexes is determined by the PEI/DNA ratio (N/P ratio i.e. number of nitrogen residues (N) in the PEI per phosphate (P) of DNA). To obtain positively charged complexes, an N/P ratio of greater than 3 is needed.
The number of nitrogen residues available in the PEI depends on the molecular weight of the polymer, its structure (branched or linear), the deprotection of the protonable residues (deacylation) and the distribution of the fragment length (polydispersity) following hydrolysis.
The linear form of PEIpro™ and the manufacturing process developed by Polyplus-transfection ensure a high, stable and reproducible amount of protonable amines available for transfection while providing a truly deacylated molecule and an extremely lower polymer chain length variation.

The quality of PEIpro™ is continuously assessed during the manufacturing process with the appropriate control testing (Fig. 1). Further, systematic lot management and release testing is performed for each lot produced. To assess activity a standardized transfection assay is performed with a suspension-adapted HEK-293 cell line in animal-component free media to ensure the reliability and reproducibility of the transfection efficiency under conditions that are suitable for biomanufacturing.

Fig. 1. PEIpro™ in-process and lot release quality controls

PEIpro™ is manufactured and formulated using a highly controlled production process (Fig. 1). Moreover, in order to meet or exceed current regulatory guidelines, PEIpro™ is released using advanced quality controls including a specification for a transfection efficiency that enables excellent lot-to-lot consistency (Fig. 2).

Fig. 2. HEK-293 EBNA cells were seeded in synthetic media, incubated at 37°C, 8% CO2 with constant shaking and transfected with PEIpro™ following the standard protocol. Luciferase expression was assayed 48 h after transfection.

PEIpro™ is free of component of animal-origin.
In addition, Polyplus-transfection is ISO 9001 Quality Management System accredited since 2002 and this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.
To ensure reliable production of recombinant proteins during the whole development process, any PEI transfection reagent selected should have the potential to meet cGMP guidelines from FDA, EMA, or ICH when the transfection reagent will be used to produce therapeutic recombinant proteins that are in late stage clinical development (ie, post Phase 1). PEIpro™ is tested to ensure complete sterility, absence of mycoplasma and very low level of bacterial endotoxins following pharmacopeia standards. In addition, on request, Polyplus-transfection can supply fully GMP compliant product delivering the highest quality level reagent and documentation for the production of therapeutic proteins or viral vectors.

Polyplus-transfection SA is the exclusive worldwide licensee of U.S. Patent No. 6,013,240, European Patent No. 0770140, and foreign equivalents, which cover transfection compositions having nucleic acids and a cationic polymer based on polyethylenimine (PEI), and use of this cationic polymer in nucleic acid transfection.
Polyplus sells branded PEI-based transfection reagents under its exclusive license, including the PEIpro™. For PEIpro™ transfection reagent sold by Polyplus and its authorized distributors, Polyplus hereby conveys license rights to the buyer to use the purchased transfection reagents, to the exclusion of human use, to perform stable or transient transfection for research and commercial purposes.
No rights to reverse engineer or resell are conveyed for any purchased transfection reagent. No license rights whatsoever are conveyed for any PEI-based molecules purchased from unauthorized sources for transfection purposes, and Polyplus reserves all rights and remedies relating to such unauthorized purchases.

Catalog Number Amount of reagent Amount of buffer 114-01 0.1 ml 5 ml 114-07 0.75 ml 60 ml 114-15 1.5 ml 2 x 60 ml 114-75 5 x 1.5 ml 120 ml x 5x conc.

1.5 ml is sufficient to perform ca.375 transfections in 6-well plates.Bulk quantities are available upon request. Please contact us.

Transfection efficiency using jetPRIME™ reaches up to 90% in cells commonly used for virus production, such as HEK-293 and derivatives, CHO, VERO and BHK cells. Hence, jetPRIME™ is the reagent for choice for viral expression vectors transfection, leading to high viral titers.

jetPRIME™ gives high viral titers for both AAV and lentivirus production.

Virus production often requires the simultaneous cotransfection of multiple plasmids to form viral particles. With its ability to efficiently compact several DNA molecules into positively charged transfection complexes, jetPRIME™ is perfectly suited for multiple plasmid cotransfection.

jetPRIME is highly efficient and only requires small amount of DNA for high transfection efficiency in adherent cells commonly used for virus production. More information on the main jetPRIME™ page.

The protocol for jetPRIME™ is easy to use since it is compatible with the use of both serum and antibiotics avoiding the need for media changes and facilitating the manipulation of a large number of vessels.

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. 4).

Figure 4. 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 complex are stable up to 8 hours at room temperature.
• 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™-HTS gives highly reproducible results at all levels:

• Well to well ► The data presented in Figure 1 were done in triplicate yielding a very small standard deviation and highly significant comparisons (Fig.1).
• One experiment to the next ► Three independent transfection experiments using the same batch of INTERFERin™-HTS show identical silencing efficiencies (Fig. 2)
• Batch to batch ► Transfection using three different batches of INTERFERin™-HTS shows highly consistent results (Fig. 3).

With INTERFERin™-HTS, your results are reliable.

Figure 2. 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 3. 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.

Catalog Number Amount of reagent Amount of buffer 114-01 0.1 ml 5 ml 114-07 0.75 ml 60 ml 114-15 1.5 ml 2 x 60 ml 114-75 5 x 1.5 ml 10 x 60 ml 114-75C 5 x 1.5 ml 120 ml 5X conc.

1.5 ml is sufficient to perform ca. 375 transfections in 6-well plates. Bulk quantities are available upon request.

Superior transfection efficiencies ranging between 70 and 90% were obtained when using jetPRIME™ reagent versus the top competitor’s reagent for several commonly used cell lines. (Fig. 1).

Fig. 1. Transfection efficiency assessed by FACS analysis in various cell lines 24 h following transfection in 24-well plates according to the manufacturer’s recommendation for competitor L2K  and 0.5 µg plasmid, 1 µl reagent per well for jetPRIME™.

Many other cell lines of various origins, as well as primary cells, are transfected with unusually high percentages (Table 1).

Table 1. Transfection efficiency of various cell types using jetPRIME. The percentage of GFP-positive cells was determined by FACS analysis 24 h after transfection.

jetPRIME™ is such a powerful in vitro transfection reagent that it only requires a small amount of reagent and plasmid DNA (Table 2), making it very economical.

Product Sheet

Table 2. Amounts of DNA and reagent (jetPRIME™ and competitor) added per well in 6-well plate for transfection according to manufacturers’ recommendations.

In addition to reducing costs, using less DNA also minimizes adverse cytotoxic effects triggered by transfection. Hence, jetPRIME™ is the reagent of choice for high transfection efficiency with excellent cell viability.

jetPRIME™ is extremely gentle to cells during transfection leading to increased cell viability (Fig. 2) and improved transfection results. Cells transfected with jetPRIME™ are healthy, while major cytotoxicity is observed with competitor.

Fig. 2. Phase contrast microscopy of HeLa cells 24 h after transfections performed according to the manufacturer’s recommendations for each reagent.

jetPRIME™ leads to over 90% knockdown of endogenous gene expression in a variety of cell lines. For example, jetPRIME™-mediated transfection of HeLa cells with 10 nM siRNA duplexes targeting endogenous lamin A/C in HeLa cells drastically reduces lamin A/C gene expression to barely detectable level (Fig. 3).

Fig. 3. Endogenous lamin A/C silencing using jetPRIME™. HeLa cells were transfected with 10 nM of 21-mer lamin A/C siRNA. After 48 h, lamin A/C silencing was assessed by immunofluorescence microscopy using an antibody against lamin A/C.

In mammalian cells, RNA interference can be achieved by other means than synthetic siRNA. Another approach consists in using plasmid-based methods. It has been reported that efficient gene silencing can be obtained with small hairpin RNA (shRNA) plasmids. For transfection of such plasmids, jetPRIME™, our versatile DNA transfection reagent, is recommended.

When studying miRNA (micro-RNA) using a plasmid-based approach, the standard transfection method using jetPRIME™ is applicable and ensures reliable transfection efficiency.

jetPRIME™ is an easy-to-use transfection reagent (Fig. 4):

• Fast and easy to scale up and down

• Compatible with serum and antibiotics

Fig. 4. jetPRIME™ convenient protocol for DNA, siRNA and co-transfection of DNA and siRNA.

Product	Catalog Number	Amount of reagent	Amount of glucose solution
in vivo jetPEI™-Gal	202-10G	0.1 ml	10 ml
in vivo jetPEI™-Man	203-10G	0.1 ml	10 ml
in vivo jetPEI™-FluoF	205-10G	0.1 ml	10 ml
in vivo jetPEI™-FluoR	206-10G	0.1 ml	10 ml

Product Catalog Number Amount of reagent jetSI™ 10 mM 403-05 0.5 ml

Please note that neither glucose solution nor DOPE are included.

1. Hassani, Z., Lemkine, G.F., Erbacher, P., Palmier, K., Alfama, G., Giovannangeli, C., Behr, J.P. and Demeneix, B.A. (2005) Lipid-mediated siRNA delivery down-regulates exogenous gene expression in the mouse brain at picomolar levels. J Gene Med 7(2): 198-207.

2. Froidevaux M. S., Berg P., Seugnet I., Decherf S., Becker N., Sachs L. M., Bilesimo P., Nygard M., Pongratz I., Demeneix B. A., (2006) The co-chaperone XAP2 is required for activation of hypothalamic thyrotropin-releasing hormone transcription in vivo EMBO Rep (7) 1182.

3. Guissouma H., Froidevaux M. S., Hassani Z., Demeneix B. A., (2006) In vivo siRNA delivery to the mouse hypothalamus confirms distinct roles of TR beta isoforms in regulating TRH transcription Neurosci Lett (406) 240-3.

4. Kumar P., Lee S. K., Shankar P. and Manjunath N. (2006) A single siRNA suppresses fatal encephalllitis induced by two different flaviviruses PLOS Med 3: 0505-1.

5. Cheret, C., Gervais, A., Lelli, A., Colin, C., Amar, L., Ravassard, P., Mallet, J., Cumano, A., Krause, K. H., Mallat, M. (2008).Neurotoxic activation of microglia is promoted by a nox1-dependent NADPH oxidase, J Neurosci 28, 12039.

6. Cakir, I., Perello, M., Lansari, O., Messier, N. J., Vaslet, C. A., Nillni, E. A. (2009) Hypothalamic Sirt1 regulates food intake in a rodent model system, PLoS One 4, e8322.

Product Catalog Number Amount of reagent Amount of 150 mM NaCl solution 112-01N 0.1 mL 5 mL FectoFly™ 112-10N 1 mL 50 mL 112-40N 4 x 1 mL 4 x 50 mL

1 ml of FectoFly™ is sufficient to perform 100 to 200 transfections in 6-well plates. For bulk sizes, Please contact us.

By simple and rapid transient transfection method, FectoFly™ provides high transfection efficiency in insect cells and robust protein production over several days (Fig. 1).

Fig 1. Protein production in S2 cells over 5 days using FectoFly transfection reagent. A suspension culture of 106 cells/ml was transfected using 2 µg pCMV-EGFPLuc and 6.4 µl of FectoFly per ml of culture in Insect-XPRESS™ medium (Lonza).

FectoFly™ provides high transfection efficiency in most commonly used insect cells (Fig. 2) allowing efficient protein production.

Figure 2. Comparison of FectoFly™ in various adherent cell lines. Sf9, Sf21, S2 or Tn5 adherent cells were transfected in 24-well plates pCMV-EGFP-Luc plasmid. Luciferase assays were performed 72 h after transfection.

FectoFly™ allows successful transfection of adherent and suspension insect cell cultures both in the latest generation of syntheticserum-free media (Fig.1, 3 – Sf21 and Sf9) and in the presence of serum (Fig.3 – S2). Transfection efficiency ranges routinely from 30% to 50% and cell viability is above 80%.

		Fig 3. GFP expression in Sf21, Sf9 and S2 cells grown in suspension, 72 h after transfection of  pCMV-EGFP-Luc plasmid using FectoFly™.

FectoFly™ was compared to other commercially available reagents dedicated to the transfection of insect cells (Fig. 4). As shown for Sf9 cells, the highest protein expression levels were obtained with FectoFly™.

Fig 4. Comparison of FectoFly™ with other insect cell-specific transfection reagents. Sf9 adherent cells were transfected in 6-well plates with pCMV-EGFPLuc plasmid using insect cell transfection reagents according to the supplier’s recommendations.

Transfection of insect cells with FectoFly™ is straightforward. o Fast : 3 steps protocol o Simple: 1 µl of FectoFly™ per µg of DNA, 1:1 ratio o Versatile: efficient in serum-free synthetic media and in the presence of serum Figure 5. FectoFly™ standard protocol.

Catalog Number Amount of reagent 115-010 10 ml 115-375 375 ml

Large sizes are available upon request. Please contact us.

PEIpro™ has been formulated at 1 mg/mL to fit current standard for transfection with PEI. This concentration eases the transfection protocol optimization. PEIpro protocol is available upon request. Contact the Technical Support.

Positively charged reagent/DNA complexes are needed to achieve high transfection efficiency. The global charge of the complexes is determined by the PEI/DNA ratio (N/P ratio i.e. number of nitrogen residues (N) in the PEI per phosphate (P) of DNA). To obtain positively charged complexes, an N/P ratio of greater than 3 is needed.
The number of nitrogen residues available in the PEI depends on the molecular weight of the polymer, its structure (branched or linear), the deprotection of the protonable residues (deacylation) and the distribution of the fragment length (polydispersity) following hydrolysis.
The linear form of PEIpro™ and the manufacturing process developed by Polyplus-transfection ensure a high, stable and reproducible amount of protonable amines available for transfection while providing a truly deacylated molecule and an extremely lower polymer chain length variation.

The quality of PEIpro™ is continuously assessed during the manufacturing process with the appropriate control testing (Fig. 1). Further, systematic lot management and release testing is performed for each lot produced. To assess activity a standardized transfection assay is performed with a suspension-adapted HEK-293 cell line in animal-component free media to ensure the reliability and reproducibility of the transfection efficiency under conditions that are suitable for biomanufacturing.

Fig. 1. PEIpro™ in-process and lot release quality controls.

PEIpro™  is manufactured and formulated using a highly controlled production process (Fig. 1). Moreover, in order to meet or exceed current regulatory guidelines, PEIpro™ is released using advanced quality controls including a specification for a transfection efficiency that enables excellent lot-to-lot consistency (Fig. 2).

Fig. 2.HEK-293 EBNA cells were seeded in synthetic media, incubated at 37°C, 8% CO2 with constant shaking and transfected with PEIpro™ following the standard protocol. Luciferase expression was assayed 48 h after transfection.

PEIpro™ is free of component of animal-origin.
In addition, Polyplus-transfection is ISO 9001 Quality Management System accredited since 2002 and this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.
To ensure reliable production of recombinant proteins during the whole development process, any PEI transfection reagent selected should have the potential to meet cGMP guidelines from FDA, EMA, or ICH when the transfection reagent will be used to produce therapeutic recombinant proteins that are in late stage clinical development (ie, post Phase 1). PEIpro™ is tested to ensure complete sterility, absence of mycoplasma and very low level of bacterial endotoxins following pharmacopeia standards. In addition, on request, Polyplus-transfection can supply fully GMP compliant product delivering the highest quality level reagent and documentation for the production of therapeutic proteins or viral vectors.

Polyplus-transfection SA  is the exclusive worldwide licensee of U.S. Patent No. 6,013,240, European Patent No. 0770140, and foreign equivalents, which cover transfection compositions having nucleic acids and a cationic polymer based on polyethylenimine (PEI), and use of this cationic polymer in nucleic acid transfection.
Polyplus sells branded PEI-based transfection reagents under its exclusive license, including the PEIpro™. For PEIpro™ transfection reagent sold by Polyplus and its authorized distributors, Polyplus hereby conveys license rights to the buyer to use the purchased transfection reagents, to the exclusion of human use, to perform stable or transient transfection for research and commercial purposes.
No rights to reverse engineer or resell are conveyed for any purchased transfection reagent. No license rights whatsoever are conveyed for any PEI-based molecules purchased from unauthorized sources for transfection purposes, and Polyplus reserves all rights and remedies relating to such unauthorized purchases.