Redian新闻
>
Re: 求助!GST fusion protein cleavage-thrombin
avatar
Re: 求助!GST fusion protein cleavage-thrombin# Biology - 生物学
t*m
1
I had the same problem once. No good way to improve it. Two pieces of
suggestion:
First, try different amount of thrombin, different incubation time and
different temperature.
Second, try different enzyme.
My problem was solved by adding a TEV cleavage site between the GST and my
target protein. TEV is a very sequence specific protease. My protein was
purified by glutathione column and eluted with TEV enzyme. It worked.
avatar
t*m
2
yes, 4oC o/n or 37oC 5,30,60 and 120min. If you do in column digestion, you
need more enzyme and your protein sample will be diluted, but you do not have
to remove the GST. To my experience, the efficiency is better. There is no
quick way to remove thrombin or GST. Any way if you want to have high quality
protein, you have to use chromatography or HPLC. In my case, after monoQ, I
got single band coomassie blue band on SDS-PAGE.

degraded
any
be
protein
相关阅读
logo
联系我们隐私协议©2024 redian.news
Redian新闻
Redian.news刊载任何文章,不代表同意其说法或描述,仅为提供更多信息,也不构成任何建议。文章信息的合法性及真实性由其作者负责,与Redian.news及其运营公司无关。欢迎投稿,如发现稿件侵权,或作者不愿在本网发表文章,请版权拥有者通知本网处理。