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Transfectamine mRNA转染试剂
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货号 | 60031 | 存储条件 | 在2-8度冷藏保存, 避免光照 |
| 规格 | 5 ml | 价格 | 11628 | |
| Ex (nm) | Em (nm) | |||
| 分子量 | 溶剂 | Water | ||
| 产品详细介绍 | ||||
简要概述
Transfectamine mRNA 转染试剂是一种功能强大且用途广泛的转染试剂，旨在将更高量的 mRNA 引入真核细胞，或者更具体地说，引入动物细胞。它在各种贴壁和悬浮细胞系（包括难以转染的细胞）中提供高转染效率。不需要核摄取，这导致比 DNA 转染更快的蛋白质表达，而没有基因组整合的风险。 Transfectamine™ mRNA 转染试剂的低毒性可提高转染细胞的活力。 Transfectamine™ mRNA 转染试剂不需要特殊培养基，与大多数商业转染试剂相比更易于使用。
产品说明书
样品实验方案
简要概述
1.准备转染细胞
2.制备Transfectamine mRNA转染试剂-RNA 混合物
3.将Transfectamine mRNA转染试剂-RNA 混合物添加到细胞培养物中
4.过夜培养细胞
5.用合适的方法分析转染效率
细胞准备
1.在转染时将细胞培养至 ~ 90% 汇合。
2.转染前更换新鲜培养基。例如，6 孔板每孔更换 2 mL 培养基，10 cm 板更换 6 mL 培养基。
工作溶液配制
1.Transfectamine mRNA转染试剂-RNA 混合物
2.将 2.5 µg mRNA 与 200 µL 无血清培养基混合。
3.在步骤 1 中加入 7.5 µL Transfectamine mRNA转染试剂。
4.混匀并在室温下孵育 20 分钟。
注意：Transfectamine mRNA转染试剂与 mRNA 的比例需要针对不同的细胞系进行优化。 通常，Transfectamine mRNA转染试剂 (µL) 与 mRNA (µg) 的比率 =（3 至 5 µL）至 1 µg。
6孔板样品方案
| 组分 | 6孔板（每孔） |
| 新鲜培养基 | 2ml |
| 纯化的mRNA | -2.5ug |
| 无血清培养基 | 200ul |
| Transfectamine mRNA转染试剂 | -7.5ul |
操作步骤
1.转染方案
1.1将Transfectamine mRNA转染试剂 – mRNA 混合物加入培养板并培养过夜。
注意：重组蛋白表达可以在转染后的8小时内检测到。 转染后约24小时可观察到最大表达水平。
图 1. HeLa 细胞中的转染效率比较（上图）和细胞毒性比较（下图）。 HeLa 细胞在 6 孔板中培养至~90% 汇合。 2.5 µg mRNA 分别用 Lipofactamin MessengerMAX 和Transfectamine mRNA转染试剂转染。 转染后 18 小时，使用带有 FITC 通道的荧光显微镜（上图）拍摄图像。 Lipofactamin MessengerMAX 和Transfectamine mRNA转染试剂的转染效率相似。用Transfectamine mRNA转染试剂转染的细胞看起来比用 Lipofatamin MessengerMAX 转染的细胞健康得多（下图）。
参考文献
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