His-tagged protein exprn in Ecoli

[Calvin and Hobbbes]

Depending on which end the label is placed(amino vs. carboxy termini) and
how the protein is folded(tertiary structure) you may or may not have
success wit this system. If the protein folds such that both ends are in
the hydrophobic interior, then the his tag will not be able to bind to the
nickel resin. This is an important note for this system to work.

Good Luck

Jon Stacks


 In article <3ijhke$fvl at newsbf02.news.aol.com>, squirenige at aol.com
(SquireNige) wrote:

> Has  anyone used the pTRCHis sert of vectors for protein expression in E
> coli, and the Qiagen Ni-NTA agarose?
> 
> Any suggestions, comments or tips would be of great help. I have just
> started trying to express a 30kDa protein, >65% GC, using SOB and
> purifying using Qiagen's NiNTA agarose, but as yet no success. 
> 
> Thanks
> 
> Nigel Walker