试剂盒提取外泌体的原理是什么

zhuyiping

最近在优化外泌体提取的方案，因为试剂盒提取量太少，超速离心对设备要求高，所以想借鉴试剂盒方法，能提取很多干细胞培养液外泌体的方法。

hereissyk

借鉴，你是想自己造一个kit吗。。你可以直接问各种kit的技术

暮色恭喜发财

我们也想自己抽提，试剂盒太贵，楼主说你们在优化提取方案，是不是已经可以抽提了？

zhuyiping

hereissyk 发表于 2015-7-31 15:03
借鉴，你是想自己造一个kit吗。。你可以直接问各种kit的技术

谢谢啦！

zhuyiping

暮色恭喜发财 发表于 2015-7-31 15:16
我们也想自己抽提，试剂盒太贵，楼主说你们在优化提取方案，是不是已经可以抽提了？ ...

很多方法都可以抽提。刚看到一篇文献，PEG6000可以沉淀，准备试试。

暮色恭喜发财

zhuyiping 发表于 2015-7-31 16:00
很多方法都可以抽提。刚看到一篇文献，PEG6000可以沉淀，准备试试。

我们也用PEG6000做过，抽提出来的粒径偏大，你们准备用什么溶液来溶解PEG，我们直接用dd水。

zhuyiping

暮色恭喜发财 发表于 2015-7-31 16:17
我们也用PEG6000做过，抽提出来的粒径偏大，你们准备用什么溶液来溶解PEG，我们直接用dd水。 ...

For preparation of 50 mL stock of 40% (w/v) PEG8000 in water, 20 grams of PEG8000 powder (Sigma) may be weighed and transferred into a 50 mL conical tube (Ambion). 20 mLs of nuclease-free water (Ambion) may then be added, and the tube placed into a Sarstedt M200 mixer (Germany) and dissolved for 1 hour or until the solution becomes clear. The contents of the tube may then be transferred into a graduated cylinder and water added to bring the final volume to 50 mL. The contents of the graduated cylinder may then be transferred back into the conical tube, and mixed for another 10 min to ensure a homogenous 40% PEG solution. Solutions containing 10-50% PEG3000-PEG20,000, with or without PBS or other buffer or NaCl, may be prepared in a similar fashion. PEG solution stocks may be stored at room temperature or 4° C. long term. Antibacterial agents such as sodium azide may be added to PEG stock solutions to prevent bacterial growth.   刚看的文献。

暮色恭喜发财

zhuyiping 发表于 2015-7-31 16:44
For preparation of 50 mL stock of 40% (w/v) PEG8000 in water, 20 grams of PEG8000 powder (Sigma) m ...

我用水提的要比用PBS的好，你提完咱们在做交流

yy8651

如果告诉你原理，试剂盒就不用卖了

chenjian00

期待结果。