超速离心法
禁止超速行驶,保持安全距离。 #生活知识# #安全常识# #交通安全法规#
2.1 外泌体纯化-超速离心法
| 2.2 外泌体纯化-密度梯度离心法
超速离心法是目前外泌体纯化中非常常用的一种方法。通过外泌体与其他细胞器的沉降系数不同,将细胞、细胞碎片以及其他细胞器等,分离出去,从而得到纯净的外泌体。
图1. 超速离心法流程示意图
△点击放大图片
1
分离细胞
首先取细胞培养液上清或体液作为样本,低速离心300 xg,10 min,分离出细胞;留上清 2000 x g离心,10 min,去除死细胞。
2
获取粗制外泌体沉淀
剩下上清继续提高离心力到10000 xg,30 min,去除细胞碎片,处理好后的上清可准备进行超速离心步骤。以100,000 xg的离心力,离心70 min,得到粗制外泌体沉淀(含有少量杂蛋白)。
3
获取纯净外泌体
用PBS重悬外泌体沉淀,再次100,000 xg,离心70 min,即可得到纯净的外泌体。
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