His-tagged proteins - bacteria or mammalian systems? - Which is better and why? (Jun/04/2006 )

I want to express and subsequently purify a Fc-myc-his tagged protein. Currently I have expressed it in a mammalian cell line and get good expression in supernatant (detected with anti-protein and anti-myc antibodies). Now I want to purify it ...

I see most people doing his-purification use bacteria ... why? Can I use mammalian system and still use nickel column kits such as His-trap FF from Amersham or those kits from Qiagen???

Please help! I am a tad clueless in this regard.

Hi Maximina,
I am in exactly in the same case as you. I expressed my protein in a mammalian cell line and I also want to purify it. Although my protein seems to not be posttranslationally modified, I was suggested to still use a mamalian cell line due to possible differences in codon translation from E. coli to mamalian cells, or other modifications. not sure of this yet
So far I tried to adapt qiagen Ni-NTA columns for the extraction of the native protein but didnt get good results. My protein elutes with the flowthrough so I am playing with imidazole concentrations and different lysis buffers. Next I will be using those his trap columns of amersham.
Lets see how we make this work out