(by novalidaddress from nurfuerspam.de)
Mon Oct 30 17:31:45 EST 2006
sometimes there's a protease site engineered before the His tag. Could
this be the case with your TEV? Then you might cut it off... But still
the His6-tag will be competing for ions on your column. And you might
dispossess devil with beelzebub by adding another protease.
Gel filtration might be the method of choice to remove your protease if
MW sizes are distant enough. Another possibility is an ion exchange
column, if your proteins have a different pI - An IEF gel should
provide you enough information.
Esperanza Fernandez wrote:
> I want to purify a his tagged protein with a ni column. The sample
> contains TEV protease since I have already performed a first
> purification step. The TEV contains a his tag so I will purify TEV
> simultaneously to my protein. Does anyone know any supplier of TEV
> protease which doesn't contain a His tag?
> Thanks a lot!
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