HiPerFect Transfection Reagent

真核细胞的siRNA和miRNA转染

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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

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Quantity

0.5 ml

1 ml

4 ml (4 x 1 ml)

100 ml

HiPerFect Transfection Reagent 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的（再）订购

特点

低浓度条件下的siRNA高效转染
高效转染原代细胞，细胞存活率高
高效转染悬浮细胞和巨噬细胞
TransFect Protocol Database中有针对特殊细胞类型的实验方案
高效转染miRNA模拟物或抑制剂

产品详情

HiPerFect Transfection Reagent是一种阳离子和中性脂组成的独特混合物，能够有效摄入siRNA和有效释放细胞内部的siRNA，即使使用低浓度的siRNA也可高效敲除基因。除了siRNA，HiPerFect Transfection Reagent还适用于低通量和高通量的转染miRNA模拟物或抑制剂。TransFect Protocol Database中有适用于特殊细胞类型的实验方案。

绩效

各种悬浮细胞系以及分化和未分化的巨噬细胞已进行过检测，通过使用HiPerFect Reagent进行siRNA转染检测。许多细胞系可成功转染，并有效敲减靶基因表达（见表1）。使用HiPerFect Reagent转染对于一些需电穿孔转染的细胞类型来说是最佳选择。

表1.使用HiPerFect Reagent成功转染的细胞系
细胞系	细胞类型	siRNA的浓度	敲减
K562	人慢性髓性白血病	5 nM	85%
Jurkat	人T细胞	75 nM	83%
D1.1	人T细胞	50 nM	84%
RAW 264.7	小鼠巨噬细胞	25 nM	77%
J774.A1	小鼠巨噬细胞	50 nM	97%
PMA-differentiated THP-1	人急性单核细胞白血病	5 nM	82%
MEL	鼠红白血病	5-20 nM	50-70%

HiPerFect Reagent不适合转染的细胞系（见表2）。

表2.HiPerFect Reagent不适合转染的细胞系
细胞系	细胞类型
Raji	人B细胞
U937	人巨噬细胞
Molt4	人T细胞
Molt14	人T细胞
HL60	早幼粒细胞系
E6.1	人T细胞

当siRNA的浓度在1–50 nM之间时，可有效敲除（>80%）（参见" HiPerFect Reagent provides effective CDC2 knockdown"）。使用1 nM siRNA转染的敲除率为86%，而5 nM siRNA转染的敲除率可达96%。最佳的siRNA使用浓度为1 nM（最少的脱靶效应并有效敲除，参见" Efficient transfection of NHEK"）或5 nM（更高的敲除效率）。

查看图表

Efficient transfection of NHEK with HiPerFect Reagent.

HiPerFect Reagent provides effective CDC2 knockdown.

原理

siRNA转染可能发生脱靶效应，即siRNAs影响非同源或部分同源靶基因的表达，但在很大程度上可通过使用低浓度的siRNA避免。使用HiPerFect Transfection Reagent，可高效的转染和沉默基因，在某些情况下可使用10 pM siRNA。使用HiPerFect Transfection Reagent可在很宽的siRNA浓度范围内高效转染siRNA，可使研究者选择他们所需的siRNA浓度。可用于转染多种类型的细胞，包括：HeLa、HeLa S3、HEK 293、NIH/3T3、Huh-7、HepG2、MCF-7、HUVEC和NHLF。

miRNA研究

micro RNAs（miRNAs）是一类内源性小分子RNA，与siRNAs特点相同。miRNAs在多种生物过程中有重要作用，如生长、分化和调亡。转染合成的miRNA模拟物和抑制剂，是一种用于解释特定miRNAs的靶标和作用的技术。miRNA模拟物是一种化学合成的miRNAs，模拟转染细胞的自然产生的miRNAs。miRNA抑制剂是一种单链修饰RNAs，能够特异性抑制miRNA的功能。转染miRNA模拟物抑制基因的表达，或转染miRNA抑制剂增强基因的表达，由此证明目的miRNA调节该基因的表达。而且，miRNAs在各种信号通路中的作用也可通过检查转染miRNA模拟物或抑制剂后特异性的基因表型来确定。筛查实验也可通过检查转染大分子量miRNA模拟物或混合物后的基因表型来进行。HiPerFect Transfection Reagent用于真核细胞siRNA的高效转染，也可用于miRNA模拟物或抑制剂的低通量或高通量转染。

程序

HiPerFect Transfection Reagent是一种即用型溶液，只需将其加入已稀释的siRNA/miRNA溶液、混合、孵育并将混合物转移到细胞即可。转染可在有血清的条件下进行，无需去除细胞中的其他物质。除了HiPerFect Transfection Reagent Handbook中的实验方案，您可以在TransFect Protocol Database中找到适合您的细胞类型和培养瓶/培养皿的实验方案。该数据库提供您所需的确切的实验方案，而不是调整已有的实验方案，节省时间和精力。只需输入细胞类型、核酸和培养瓶的类型即可打印或下载QIAGEN转染实验方案的PDF格式。TransFect Protocol Database是免费的，且无需注册。

HiPerFect Transfection Reagent转染，可使用常规的转染步骤，在转染前24小时种细胞。同时也提供使用96孔板和384孔板反转染的实验方案（见HiPerFect Transfection Reagent Handbook）。这些实验方案，可在同一天种细胞和进行转染。加入HiPerFect Reagent后，将siRNA/miRNA加入孔中。混合均匀后，将细胞加入孔中（参见" Reverse transfection using HiPerFect Transfection Reagent"）。反转染实验方案快速、方便、可实现自动化，通常用于高通量实验。

查看图表

Reverse transfection using HiPerFect Transfection Reagent.

应用

即使siRNA浓度较低，HiPerFect Transfection Reagent也能够高效进行siRNA转染，适合多种应用，如：

RNA干扰研究
miRNA研究
基因表达和功能研究

辅助数据和图表

Efficient transfection of NHEK with HiPerFect 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.

Specifications

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

资源

Brochure detailing reagents for efficient and robust DNA and RNA transfection.

Poster for download

For transfection of eukaryotic cells with siRNA and miRNA

Download Safety Data Sheets for QIAGEN product components.

FAQ

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

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

FAQ ID -972

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

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.

-1

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

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

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

After transfection, remove the medium from the cells and wash the cells once with PBS.
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)
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

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

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

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

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

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

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

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.

http://www.sabiosciences.com/reversetransfection.php

FAQ ID -2777

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