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 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.

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Product Cat. no. List price:
HiPerFect Transfection Reagent (0.5 ml)
HiPerFect Transfection Reagent for up to 166 transfections in 24-well plates or up to 666 transfections in 96-well plates
301704
$204.00
HiPerFect Transfection Reagent (1 ml)
HiPerFect Transfection Reagent for up to 333 transfections in 24-well plates or up to 1333 transfections in 96-well plates
301705
$358.00
HiPerFect Transfection Reagent (4 x 1 ml)
HiPerFect Transfection Reagent for up to 1332 transfections in 24-well plates or up to 5333 transfections in 96-well plates
301707
$1,262.00
HiPerFect Transfection Reagent (100 ml)
HiPerFect Transfection Reagent for transfections in up to 1388 96-well plates
301709
$25,750.00
The HiPerFect Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Efficient transfection of NHEK with HiPerFect Reagent.|HiPerFect Reagent provides effective CDC2 knockdown.|Reverse transfection using HiPerFect Transfection Reagent.|
Normal human embryonal keratinocytes were transfected with siRNA targeting lamin A/C or with AllStars Negative Control siRNA. Lamin A/C expression was measured by real-time RT-PCR after 48 hours.|HeLa S3 cells were transfected with a range of concentrations of siRNA targeted against CDC2 using HiPerFect Transfection Reagent from QIAGEN or Reagent L from another supplier. Non-silencing siRNA targeting green fluorescent protein (GFP) was also transfected. After 48 hours, CDC2 knockdown was assessed using quantitative, real-time RT-PCR. CDC2 expression was normalized to the expression of GAPDH. Expression levels relative to untransfected control cells are shown.|In reverse transfection 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. These transfection protocols are rapid and convenient, can easily be automated, and are frequently used for high-throughput experiments.|
Performance

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).

Principle

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.

Procedure

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.

Applications

 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
Feature
Specifications
Applications RNAi studies, gene expression studies
Cell type Eukaryotic cells including primary cells
Controls Not included
Features Minimized risk of off-target effects, rapid reverse transfection
Nucleic acid siRNA, miRNA
Number of possible transfections 333 transfections in 24-well plates / 1 ml reagent
Technology Cationic and neutral lipids
Transfection type Transient transfection

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