HiPerFect Transfection Reagent

For transfection of eukaryotic cells with siRNA and miRNA


  • Efficient transfection using low siRNA concentrations
  • Effective transfection of primary cells with high cell viability
  • Effective transfection of suspension cells and macrophages
  • Cell-specific protocols at the TransFect Protocol Database
  • Efficient transfection of miRNA mimics or inhibitors
HiPerFect Transfection Reagent (0.5 ml)

Cat. No. / ID: 301704

HiPerFect Transfection Reagent for up to 166 transfections in 24-well plates or up to 666 transfections in 96-well plates
0.5 ml
4 ml (4 x 1 ml)
1 ml
100 ml
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The HiPerFect Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

HiPerFect Transfection Reagent is a unique blend of cationic and neutral lipids that enables effective siRNA uptake and efficient release of siRNA inside cells, resulting in high gene knockdown even when using low siRNA concentrations. In addition to siRNA, HiPerFect Transfection Reagent is ideally suited to transfection of miRNA mimics or inhibitors. Cell-type-specific protocols using HiPerFect Transfection Reagent are available at the TransFect Protocol Database.


A range of suspension cell lines and differentiated and undifferentiated macrophages have been tested using HiPerFect Reagent for siRNA transfection.  Many lines were successfully transfected with excellent knockdown of target gene expression (see Table 1). Transfection with HiPerFect Reagent is a superior alternative to electroporation for many of these cell types. 

Table 1. Cell lines successfully transfected using HiPerFect Reagent protocols
Cell line Cell type siRNA concentration Knockdown
K562 Human chronic myeloid leukemia 5 nM 85%
Jurkat Human T-cell 75 nM 83%
D1.1 Human T-cell 50 nM 84%
RAW 264.7 Mouse macrophage 25 nM 77%
J774.A1 Mouse macrophage 50 nM 97%
PMA-differentiated THP-1 Human acute monocytic leukemia 5 nM 82%
MEL Murine erythroleukemia 5-20 nM 50-70%

Some cell lines were not amenable to transfection using HiPerFect Reagent (see Table 2). 

Table 2. Cell lines not amenable to transfection using HiPerFect Reagent protocols
Cell line Cell type
Raji Human B-cell
U937 Human macrophage
Molt4 Human T-cell
Molt14 Human T-cell
HL60 Promyelocytic cell line
E6.1 Human T-cell

Efficient knockdown (>80%) is achieved with siRNA concentrations in the range of 1–50 nM (see figure " HiPerFect Reagent provides effective CDC2 knockdown"). Transfection of 1 nM siRNA resulted in 86% knockdown and transfection of 5 nM siRNA increased the knockdown efficiency to 96%. Depending on the purpose of the RNAi experiment, the optimal concentration of siRNA to use may be 1 nM (minimal risk of off-target effects and efficient knockdown) or 5 nM (higher knockdown efficiency).

See figures


Transfection of siRNA can result in off-target effects, in which siRNAs affect the expression of non-homologous or partially homologous gene targets. Off-target effects, which may produce misleading results in RNAi experiments, can be largely avoided by using low siRNA concentrations. Using HiPerFect Transfection Reagent, highly efficient transfection and silencing have been observed, in some cases with as little as 10 pM siRNA. HiPerFect Transfection Reagent provides highly efficient siRNA transfection over a range of siRNA concentrations from low to high, allowing researchers to choose the siRNA concentration they wish to use. It can be used to transfect a wide range of cell types, including HeLa, HeLa S3, HEK 293, NIH/3T3, Huh-7, HepG2, MCF-7, HUVEC, and NHLF (see figure "Efficient transfection of NHEK").

miRNA research

microRNAs (miRNAs) are a class of endogenous small RNA molecules with similar characteristics to siRNAs. miRNAs play a role in many diverse biological processes such as development, differentiation, and apoptosis. Transfection of synthetic miRNA mimics or inhibitors is a technique used to elucidate the targets and roles of particular miRNAs. miRNA mimics are chemically synthesized miRNAs which mimic naturally ocurring miRNAs upon transfection into the cell. miRNA inhibitors are single-stranded modified RNAs which specifically inhibit miRNA function. Reduced gene expression after transfection of an miRNA mimic, or increased expression after transfection of an miRNA inhibitor, provide evidence that the targeted miRNA is involved in regulation of that gene. Alternatively, the role of miRNAs in various pathways can be studied by examination of a specific phenotype after transfection of an miRNA mimic or inhibitor. HiPerFect Transfection Reagent was developed for highly efficient transfection of eukaryotic cells with siRNA, and is ideally suited to transfection of miRNA mimics or inhibitors.


HiPerFect Transfection Reagent is provided as a ready-to-use solution - just add the reagent to your diluted siRNA/miRNA, mix, incubate, and pipet the complexes onto the cells. Transfections can be performed in the presence of serum, eliminating the need to remove complexes from the cells. In addition to the protocols provided in the HiPerFect Transfection Reagent Handbook, you can find protocols to suit your cell type and plate/dish format using the TransFect Protocol Database. The database takes the guesswork out of transfection protocols. Rather than adapting existing protocols to fit your requirements, the database provides exactly the protocol needed, saving time and effort. Simply enter the cell type, nucleic acid, and plate format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format. Use of the TransFect Protocol Database is free of charge and no registration is required.

Transfection with HiPerFect Transfection Reagent can be carried out using the common transfection procedure of seeding the cells 24 hours before transfection. Alternatively, protocols are provided for reverse transfection in 96-well plates and 384-well plates (see the HiPerFect Transfection Reagent Handbook). In these protocols, cells are seeded and transfected in the same day. siRNA/miRNA is spotted into wells followed by the addition of HiPerFect Reagent. After complex formation, cells are added to the wells (see flowchart " Reverse transfection using HiPerFect Transfection Reagent"). Reverse transfection protocols are rapid, convenient, and can easily be automated.

See figures


 HiPerFect Transfection Reagent enables highly efficient siRNA transfection, even with low siRNA concentrations, for applications such as:

  • RNA interference studies
  • miRNA research
  • Studies on gene expression and function

Supporting data and figures


ApplicationsRNAi studies, gene expression studies
Transfection typeTransient transfection
FeaturesMinimized risk of off-target effects, rapid reverse transfection
Nucleic acidsiRNA, miRNA
TechnologyCationic and neutral lipids
ControlsNot included
Cell typeEukaryotic cells including primary cells
Number of possible transfections333 transfections in 24-well plates / 1 ml reagent


Brochures & Guides (5)
Scientific Posters (1)
Poster for download
Kit Handbooks (2)
For transfection of eukaryotic cells with siRNA and miRNA
Quick-Start Protocols (1)


Which transfection reagent is recommended for transfection of the miScript Target Protectors?
HiPerfect Transfection Reagent is recommended for the transfection of the miScript Target Protector alone. Attractene Transfection Reagent is recommended for DNA/oligo co-transfection.
FAQ ID -2252
Do you have a protocol for long-term, siRNA-mediated gene silencing using HiPerFect Transfection Reagent?

Yes, please follow the Protocol 'Long-Term, siRNA-Mediated Gene Silencing' in the HiPerFect Transfection Reagent Handbook.


FAQ ID -972
Do you have transfection data for QIAGEN Transfection Reagents?

QIAGEN Transfection Reagents have been used successfully with many different cell types. For your convenience, we have organized data from researchers who have shared their experimental results with us online in our Transfection Cell Database. Simply type your cell line of interest into the 'Search' field on this page, and find transfection results achieved with various QIAGEN Transfection Reagents.  Please note that QIAGEN cannot verify data supplied from outside sources.

You can also submit your own transfection data and obtain a gift as appreciation.  In addition you can find on our TransFect Protocol Database transfection protocols for specific cell types and plate formats.

FAQ ID -158
Do you have a protocol for transfection of differentiated macrophage cell lines (THP) with siRNA?

Yes, please follow the protocol "Transfection of Differentiated Macrophage Cell Lines, Including THP-1, with siRNA in 24-Well Plates" in the HiPerFect Transfection Reagent Handbook.

How many transfections can I perform with 20 nmol of HPP grade siRNA?

The number of transfections you can perform with 20 nmol HPP grade siRNA depends on the plate format used, the Transfection Reagent, and additional optimization requirements for your specific application.

In general, when using the HiPerFect Transfection Reagent in a 24-well plate format with 37.5 ng of siRNA per transfection you should be able to perform at least 6600 transfections.

If you are interested in other siRNA synthesis scales and purification options, please visit the QIAGEN Custom siRNA Synthesis page and our GeneGlobe data base, where you can find a large selection of pre-designed and validated siRNAs targeted at human, mouse and rat genes.

FAQ ID -367
What is the difference between HiPerFect and HiPerFect HTS?

The composition is confidential.   HTS is a different component from HiPerFect.  HTS works with very low amounts of reagent for transfection, and is specifically adapted for use in high throughput applications.  HiPerFect Transfection Reagent is for siRNA transfection at lower throughput applications.

FAQ ID -3108
Do you have a protocol for the fixation of cells transfected with fluorescently labeled siRNA?

For the fixation of cells transfected with fluorescently labeled siRNA, we would suggest to perform the following protocol:

  1. After transfection, remove the medium from the cells and wash the cells once with PBS.
  2. Incubate the cells for 15 minutes at room temperature with 4% Paraformaldehyde (in PBS, pH 7.0). The cells should be completely covered by this solution (e.g., for a 96-well plate use 50 µl solution/well)
  3. Wash the cells with PBS.

Fixed cells can be stored at 4°C for a few days.

(Note: It is also possible to use chamber slides or object slides for this procedure. Object slides should be coated to provide better growing conditions for cells. Cells can be fixed as described above and then overlayed with embedding medium to allow investigation using a fluorescence microscope. Optimal conditions for this method need to be determined by the user)


FAQ ID -793
Do you have a protocol for transfection of suspension cell lines (Jurkat and K562) with siRNA?

Yes, please follow the protocol "Transfection of Suspension Cell Lines, including Jurkat and K562, with siRNA in 24-Well Plates" in the HiPerFect Transfection Reagent Handbook.



FAQ ID -1250
How can I transfect siRNA into insect cells such as Drosophila melanogaster S2?

Unfortunately, we do not have any data for the transfection of siRNA into insect cells. However, we know about customers who have successfully used Effectene Transfection Reagent for the transfection of plasmid DNA into insect cell lines S2 and Sf9 (see FAQ 397). Since Effectene Transfection Reagent and HiPerFect Transfection Reagent for the transfection of eukaryotic cells with siRNA are both lipid based, there is a chance that HiPerFect will work with insect cells. It is especially suitable for transfection of very low siRNA concentrations, reducing the risk of cell toxicity and off-target knockdown effects.

Alternatively, you can try RNAifect or TransMessenger Transfection Reagent, also lipid-based, for the transfection of siRNA into S2 cells.

We would appreciate any feedback on your results in case you want to give it a try. Please visit our Transfection Cell Survey page to submit your feedback.

FAQ ID -1046
Do you have a protocol for transfection of macrophage cell lines (J774.A1 and RAW 264.7) with siRNA?

Yes, please follow the protocol "Transfection of Macrophage Cell Lines, Including J774.A1 and RAW 264.7, with siRNA in 24-Well Plates" in the HiPerFect Transfection Reagent Handbook.



FAQ ID -1251
How much dye-labeled siRNA should I use to monitor transfection efficiency when using HiPerfect Transfection Reagent?
The amount of HiPerfect Transfection Reagent and siRNA required for optimal performance may vary, depending on the cell line and gene target. When transfecting fluorescently labeled siRNA (e.g., Alexa Fluor 488 siRNA) for monitoring transfection efficiency under a fluorescent microscope, it may be necessary to increase the amount of siRNA up to 25 nM (without increasing the volume of HiPerFect Reagent), depending on the optical requirements of the microscopic equipment that will be used for detection.
FAQ ID -1058
Do you have a protocol for reverse transfection of adherent cells with siRNA in 384-well plates?

Yes, please follow the protocol 'Reverse Transfection of Adherent Cells with siRNA in 384-Well Plates' in the HiPerFect Transfection Reagent Handbook.


FAQ ID -971
Does HiPerFect work for the transfection of miScript miRNA mimics and inhibitors?

HiPerFect Transfection Reagent does work for the transfection of miScript miRNA Mimics and Inhibitors, and the new handbook contains protocols for this application.



FAQ ID -1984
What is the most reliable transfection reagent for delivering shRNA plasmids and siRNA to cells in culture?

HiPerFect Transfection Reagent is optimized for siRNA transfections and enables effective siRNA uptake and efficient release of siRNA inside cells, resulting in high gene knockdown even when using low siRNA concentrations. For high-throughput siRNA screenings HiPerFect HTS Reagent is a fast and effective newcomer specifically designed to focus on robustness and cost efficiency. For the transfection of shRNA.

Attractene Transfection Reagent should be used for the transfection of shRNA (short-hairpin RNA) vectors for gene silencing experiments to achieve high efficiency. Ease and flexibility of handling enables preparation and storage of transfection complexes making Attractene Reagent suitable for use with automated systems.


FAQ ID -2777
What is the average molecular weight of a siRNA, and how do I convert uM to ug values?
The Molecular Weight (MW) of a 21 nucleotide double-stranded siRNA molecule is approximately 13-15 ug/nmol. The exact MW depends on the sequence of the siRNA. 20 uM of double-stranded 21 nt siRNA is equivalent to approximately 0.25 ug/ul.
FAQ ID -388