产品说明书
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已发表文献
产品描述
Polybrene（聚凝胺，hexadimethrine bromide）是一种多聚阳离子聚合物，常用于哺乳动物细胞的DNA转染实验以增强脂质体的转染效率。Polybrene目前广泛用于逆转录病毒介导的基因转染，慢病毒介导的基因转染，作用机理可能是通过中和细胞表面唾液酸与病毒颗粒之间的静电排斥从而促进吸附作用。Polybrene也是一种有名的抗肝素剂（肝素拮抗剂），常用来生产非特异性凝集的红细胞。另外Polybrene也多用于蛋白测序，因为小剂量的Polybrene在自动测序分析可明显改善多肽的降解现象。PVDF膜加入polybrene还能提高膜的亲和性。
本产品以溶液形式提供，粉末用0.9% NaCl配制成10 mg/mL的溶液，并用0.22 μM滤膜过滤除菌。使用时一般按1:1000-1:2000稀释，依细胞种类不同稀释比例不同，具体查阅相关文献。
运输与保存方法
冰袋（wet ice）运输。-20 ºC可保存2年，建议分装保存，避免反复冻融。
注意事项
1）为了您的安全和健康，请穿实验服并戴一次性手套操作。
2）本产品仅作科研用途！
操作步骤(仅供参考，具体使用浓度请参考相关文献)
【注】：Polybrene对某些细胞（如末端分化的神经元，DC细胞）毒性较大，初次应用建议先做毒性测试。
实验1：逆转录病毒感染（Retroviral Infection）
（1）重组逆转录病毒原液的制备：取5 mL生长培养基（5%血清）加入含单层转染逆转录包装细胞的100 mm培养盘内。孵育24 h后，吸去培养液并用0.45 μm滤器过滤。
（2）待感染细胞的培养：100 mm培养盘内加入10 mL完全培养基，细胞密度为5×105/盘。
（3）病毒感染：细胞培养24 h后，吸去完全培养液。用含polybrene的2 mL病毒上清 （或将病毒原液稀释到2 mL）感染细胞，polybrene的终浓度为5-10 μg/mL。37°C孵育3-6 h。
（4）收集病毒颗粒：加入8 mL完全培养基。培养2-3天后，收集培养基上清，处理得到病毒颗粒。
实验2：转染
（1）完全生长培养基培养细胞，培养细胞密度约50%；
（2）孵育细胞18-24 h后准备DNA-培养基- Polybrene混合液，按如下操作制备混合液：①添加完全培养基（60 mm培养皿2 mL，100 mm培养皿3 mL），37°C预热；②添加10 ng~10 µg质粒轻轻混匀；③加入Polybrene至终浓度为5-10 μg/mL。轻轻混匀。以上每个成分需要按顺序加入。
（3）去除培养基，在细胞中加入DNA-培养基-Polybrene溶液，在37°C孵育细胞6-20 h。细胞培养的前6 h内约每1.5 h轻柔混匀。
Ver.CN20240805
Q: 聚凝胺浓度是10mg/ml，用什么稀释？
A: 聚凝胺稀释不是指用其他溶液稀释，而是指在具体实验中加入的比例，比如实验体系有10mL,按照这个比例（1：1000为例），就直接取10uL的产品（10mg/mL溶液）加入实验体系里面即可。
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