血清外泌体抽提方法y

暮色恭喜发财

除了超速离心和密度梯度离心外，就只能用试剂盒提取了，大家有谁知道试剂盒提取的原理是什么呢，用哪些试剂可以代替试剂盒呢，即使产量跟纯度差一点也没关系。

chyi

顶，我也想知道

惜名

所有方法在这张帖子都有罗列：http://www.exosome.com.cn/forum. ... p;extra=&page=1
题主，life、SBI、101bio的试剂盒我们都算过价格了，都不便宜。。。
这个课题目前就是这样烧钱的状态，超离得率低，需要耗费大量培养基
如果贵实验室这点钱都无法支撑，我劝还是赶紧转方向吧

暮色恭喜发财

Johnny 发表于 2015-8-4 18:01
一哥常逛我们论坛哈

欢迎欢迎

当然是降低成本了，不受试剂盒的限制。

chyi

Johnny 发表于 2015-8-4 18:01
一哥常逛我们论坛哈

欢迎欢迎

  觉得问题不错，顶一下

zhuyiping

  1.For the isolation of exosomes from human umbilical cord Wharton’s jelly-MSCs and human dermal fibroblasts, serum-free culture medium conditioned for 24 hours was filtered (0.2 m) and concentrated by ultrafiltration device with 100-kDa cutoff (Millipore).
   2.Exosomes in CM were precipitated with one-third volume of polyethylene glycol buffer (33.4% PEG 4000, 50 mmol/L HEPES [pH 7.4], 1 mol/L NaCl) overnight at 4°C, followed by centrifugation at 12 000 g for 5 minutes and resuspension in PBS (pH 7.4).
   3.Exosomes in polyethylene glycol–precipitated fraction were further purified by S200 size-exclusion chromatography. A 75-L sample was applied on a S200 column (Clontech, Mountain View, CA)pre-equilibrated with PBS by spinning at 700 g for 5 minutes, and the exosomal fraction was subsequently eluted in the flow-through by
centrifugation at 700 g for 5 minutes.刚看的文献。

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zhuyiping 发表于 2015-8-5 16:07
1.For the isolation of exosomes from human umbilical cord Wharton’s jelly-MSCs and human dermal f ...

非常感谢，明天我试试，看看效果。

暮色恭喜发财

zhuyiping 发表于 2015-8-5 16:07
1.For the isolation of exosomes from human umbilical cord Wharton’s jelly-MSCs and human dermal f ...

这篇文献能发我一份吗？464355399@qq.com

mirexosome

能给我一份吗

1.For the isolation of exosomes from human umbilical cord Wharton’s jelly-MSCs and human dermal f ...

，1040269617@qq.com,
发财楼主效果如何之后跟我们分享一下，谢谢

暮色恭喜发财

一定，相互学习，共同进步。