Purification of HisTag Recombinant Protein - Advices needed (Aug/06/2007 )

I cloned my protein together with HisTag and want to purify it.
I used pQE series (Qiagen) and their range of products are just massive and I am just pretty much confused.

The Ni NTA Matrices are:

NiNTA Agarose
NiNTA Superflow
NiNTA spin column

I bought the spin column and realise it is just for small scale.

I am trying to purify in a large scale and in native condition.
Which one should I get? the Agarose or superflow?
What are the differences?

And also.. i think I will need some polypropylene columns as well right?

Thank you so much for all the inputs (if there is).

Hi,

In short, the Agarose is for our normal protein purification application (bind proteins to the beads, wash through column, elute...), I think they call that as gravity flow. The Superflow is more useful for big scale production and FPLC. You can check more information here: http://www1.qiagen.com/Products/Protein/Purification.aspx

About the column, I think that you did purify GST proteins before? We did not use different kind of column for His-tagged and GST tagged in our lab.

hello there,

thanks for the reply. I had checked. I think Superflow is suitable for FPLC.

I read one of the reviews saying that the polypropylene tube is packed with NiNTA. So I am not really sure how the system works now. Is it with Ni NTA or without?

Argh.. protein purification is driving me nuts.