HiPerFect Transfection Reagent (4x1 ml)，301707，Qiagen，凯杰

HiPerFect Transfection Reagent

真核细胞的siRNA和miRNA转染

Features

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

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。

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

HiPerFect Reagent不适合转染的细胞系（见表2）。

  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）。

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.

Reverse transfection using HiPerFect Transfection Reagent.

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.

HiPerFect Reagent provides effective CDC2 knockdown.

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.

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