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Biology版 - how to purify the degraded protein bands on the gel?
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相关话题的讨论汇总
话题: protein话题: degraded话题: bands话题: gel话题: purify
进入Biology版参与讨论
1 (共1页)
c*******4
发帖数: 41
1
My protein is around 20kDa, and I see many degraded protein bands (from
10kDa to 3kDa) on the gel. I tried to wash my protein several times using
the amicon centrifugal filter at 10kDa cutoff, but I still see those
degraded protein bands on the gels. I also add proteinase inhibitor and also
do the purification on ice to prevent the degradation, but it does not help
. Does anyone know what is the best way to purify or get ride of these
degraded protein bands on the gel? thanks.
e****s
发帖数: 1125
2
Your title is really confusing.
This could be very challenging. Because the truncated forms, especially
those with similar size to the full-length, behave very similar to full-
length protein. If most of your truncated is smaller than half of the full-
length, then it is doable. Gel filtration may helps to remove those big
truncated ones. You can also try those traditional columns, such as ion-
exchanged or hydrophobic columns.
Amacon's concentrator was not designed to do this.
I will suggest to prevent it from happening by adding protease inhibitors;
however,sometimes it is just the way it is.
If you really need full-length proteins, you can also add different tags at
each side.

also
help

【在 c*******4 的大作中提到】
: My protein is around 20kDa, and I see many degraded protein bands (from
: 10kDa to 3kDa) on the gel. I tried to wash my protein several times using
: the amicon centrifugal filter at 10kDa cutoff, but I still see those
: degraded protein bands on the gels. I also add proteinase inhibitor and also
: do the purification on ice to prevent the degradation, but it does not help
: . Does anyone know what is the best way to purify or get ride of these
: degraded protein bands on the gel? thanks.

1 (共1页)
进入Biology版参与讨论
相关主题
Please recommend good Glut1 and Glut4 antibodies.请教一个拉丁用法
why size is so big different?发现一个protein interaction
questions about mMESSENGERmMACHINE sp6请问有没有什么公司purify untagged recombinant protein from human cells?
大家有碰到过Co-IP突然不work的情况吗?请问一下为啥要在4度房子里玩蛋白?
请推荐protein buffer exchange columnhow to store cell lysate for a long time
How to test protein stabilitypostdoc position on membrane protein purification
Amicon ultra-15 centrifugal filter 穿滤公司订制的抗体在wild type和knock out 小鼠都检测到band了, 是小鼠问题还是抗体问题?
请教几个用来产生 membrane proteins设备的价格 (转载)【请问】有commercially available的purified Cas9 Protein 吗?
相关话题的讨论汇总
话题: protein话题: degraded话题: bands话题: gel话题: purify