想统计下大家用超速离心时一次用多少量的细胞上清才够用

木瓜31

luckkit 发表于 2015-4-24 12:38
另外，请教一下，俺是收集上清后直接2000g 30min,然后0.22um过滤，10e4 30min, 10e5 2hrs, 开始没有300g 10 ...

300g pellet=cell,2000g pellet=dead cell.个人感觉先以较低速度把细胞离下比较合理，也许直接上2000g细胞应激下会分泌外泌体？ 谁知道呢。。

bobo

woshiyywlly 发表于 2015-4-21 13:48
对照和实验组各16个dish

实验组和对照组怎么设置啊？

ashey

木瓜31 发表于 2015-5-7 21:43
300g pellet=cell,2000g pellet=dead cell.个人感觉先以较低速度把细胞离下比较合理，也许直接上2000g细 ...

可以再了解一下各个离心条件下离出来的沉淀和上清分别是什么。

木瓜31

D:\桌面陈雅静\外泌体之家\QQ图片20150525163753.png

木瓜31

ashey 发表于 2015-5-21 08:30
可以再了解一下各个离心条件下离出来的沉淀和上清分别是什么。

不会发图。。你看看这篇文章吧Isolation and Characterization  of  Exosomes from Cell Culture Supernatants
and Biological Fluids

bobo

木瓜31 发表于 2015-5-25 16:40
不会发图。。你看看这篇文章吧Isolation and Characterization  of  Exosomes from Cell Culture Superna ...

我想用试剂盒提exosome,那么如何将100ml的细胞培养液浓缩到4ml,从而使培养液中的exosome浓度大些

woshiyywlly

bobo 发表于 2015-5-20 23:24
实验组和对照组怎么设置啊？

这个要看你实验要求的

evomicscience

luckkit 发表于 2015-4-24 12:38
另外，请教一下，俺是收集上清后直接2000g 30min,然后0.22um过滤，10e4 30min, 10e5 2hrs, 开始没有300g 10 ...

the 1st 2000g may damage cell which then release miRNAs and/or proteins etc. You should use 300g to pellet cells,  and then 2000g for apoptotic bodies, 0.22um or 10k for microvesicle.

seallin

evomicscience 发表于 2015-6-2 12:55
the 1st 2000g may damage cell which then release miRNAs and/or proteins etc. You should use 300g t ...

What is "0.22um or 10k for microvesicle" mean? 0.22 is the flow-through? And 10K is for concentration?

seallin

evomicscience 发表于 2015-6-2 12:55
the 1st 2000g may damage cell which then release miRNAs and/or proteins etc. You should use 300g t ...

What is "0.22um or 10k for microvesicle" mean? 0.22 is the flow-through? And 10K is for concentration?