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TransMessenger Transfection Reagent

高效的mRNA转染

  • 高效的转染
  • 严格的质量监控确保可靠的结果
  • 高效转染原代细胞

TransMessenger Transfection Reagent是即用型、脂质体试剂,用于真核细胞的RNA转染。使用RNA而非DNA转染的细胞为转染实验提供了新的可能性。使用TransMessenger Reagent成功转染的细胞株包含293T、Jurkat细胞和Vero细胞。该产品也可用于RNA转染人类原代成纤维细胞、大鼠心肌细胞和大鼠神经细胞。

Cat No./ID: 301525
TransMessenger Transfection Reagent (0.5 ml)
For 60 transfections in 6-well plates or 80 transfections in 12-well plates

TransMessenger Transfection Reagent适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
RNA与TransMessenger Reagent的量及其转染效率。
用CHO-K1细胞进行优化实验。将2 x 104个细胞一式四份接种到96孔板中,24小时后用体外转录的CAT编码RNA转录。[A]使用加量的RNA和1.5 µl TransMessenger Reagent,[B]使用加量的TransMessenger Reagent和0.25 µg RNA,按照TransMessenger Transfection Reagent操作手册进行实验。转染后24小时检测CAT活性。
1
TransMessenger Reagent和HeLa细胞。
检测HeLa-S3细胞中的绿色荧光蛋白(GFP)。将8 x 104个细胞接种到48孔板中,24小时后用0.5 µg体外转录的GFP编码RNA(转录自P7ASP-GFP/Mlu)、1 µl Enhancer R和2.5 µl TransMessenger Reagent进行转录。转录24小时后,通过荧光显微镜分析细胞。约有50%的细胞被成功转染。(P7ASP-GFP/Mlu为美国斯坦福大学血液中心J. Bogenberger提供)
2
Cardiac cell studies.
[A] Use of TransMessenger Transfection Reagent resulted in an excellent, 60% survival rate post-transfection with beating, functional cardiomyocytes, and a transfection efficiency of 10–13%. [B] High rate of cardiomyocyte and non-cardiomyocyte transfection. Left: Superimposed images showing anti-GFP (green; transfected cell) and anti-α-actinin (red; cardiomyocyte) staining. Right: Anti-α-actinin (cardiomyocyte) staining. Arrows = GFP-positive cardiomyocytes;* = GFP-positive non-cardiomyocytes. [C]  Left: GFP-positive cardiomyocytes (green) and non-transfected cardiomyocytes (red; stained with anti-α-actinin) demonstrate equivalent morphologies. Right: Transfected cardiomyocyte and non-cardiomyocyte cells.
Data provided by QBMCellScience Inc, Ottawa, ON. Canada
Performance
TransMessenger Reagent提供快速简单的操作流程和高转染效率(参见"TransMessenger Reagent with HeLa cells")。因为RNA量是成功转染的关键因素,我们推荐对于每个细胞类型和RNA,都优化RNA和TransMessenger Transfection Reagent量(参见"Amount of RNA and TransMessenger Reagent vs. transfection efficiency")。为了操作便利,该试剂配套提供优化准则和在不同细胞培养物规格下的优化起始点。
Principle

TransMessenger Transfection Reagent是首款专为用RNA转染细胞的试剂。该试剂由脂质体制备,与特定的RNA冷凝增强剂和优化的缓冲液一起使用。RNA分子通过增强剂冷凝,然后涂上TransMessenger Reagent有效地转移到真核细胞中。严格的质量监控用于测试无RNA酶活性、每批次的一致性和低内毒素水平(≤10 EU/ml)。我们严格的标准消除了试剂的变量,避免对RNA转染效率产生不利影响。

Procedure

TransMessenger Reagent为即用型试剂。为获得TransMessenger–RNA转染混合物,只需简单混合具增强剂R的RNA和Buffer EC-R,并在室温下孵育5分钟,然后加入TransMessenger Reagent再孵育5–10分钟。将其与培养液混合并直接加入细胞中。随后需要3小时的孵育,培养液发生改变,孵育细胞直到可用于分析应用。使用高纯度的RNA可获得最佳的转染结果,无DNA、蛋白质和其他污染物。推荐使用RNeasy和Oligotex mRNA Kits纯化RNA。

Applications

RNA而非DNA转染的细胞为转染实验提供新的可能性。转染RNA序列的表达无需转录、不依赖于启动子。此外,蛋白表达通常会在RNA而非DNA转染后快速出现。使用TransMessenger Transfection Reagent的RNA转染可用于:

研究无法用质粒DNA高效转染的细胞
直接研究RNA功能
Features
Specifications
应用 Direct studies of RNA function
细胞类型 Eukaryotic cells
对照 Not included
特点 Transfection with RNA. Efficient transfection of neuronal cells. Tested for the absence of RNases.
核酸 RNA
可转染的次数 80 transfections in 12-well plates / 0,5 ml reagent
技术 Lipid-based formulation in conjunction with a RNA-condensing enhancer
转染类型 Transient transfection, co-transfection

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产品介绍与指南 (2)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
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试剂盒操作手册 (1)
For transfection of eukaryotic cells with RNA and siRNA
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转染实验方案 (2)
Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.
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Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.
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补充实验方案 (1)
The following procedure is for co-transfection of adherent cells using siRNA and plasmid DNA in one well of a 24-well plate. This procedure is provided as a starting point for optimization of siRNA and plasmid DNA co-transfection in mammalian cells using TransMessenger™ Transfection Reagent.
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