Hello all,
In my research I intend to purify ChlG (a membrane protein) from cyanobacteria using a protein tag. I want to elute the protein together with its interacting partners or complexes. I have read that the Tandem Affinity Purification (TAP) tag is a method suited for this particular purpose. Nevertheless, I did't understand how is it better than other tags, such as Flag or Rho1D4, in the purification of complexes. From what I understood, the only difference is that in TAP there are two seperate steps of affinity purification.
Also, I wanted to ask whether this method is at all compatible with cyanobacteria?
Thanks very much,
Or
TAP tags are good but potentially not for anything with delicate metal binding sites.
The Calmodulin Binding Protein component is eluted with EGTA.
In your case you are looking to pull down interacting partners, you never know what that might do to your complex.
So alternative TAP combinations might be worthwhile for you, substituting the CBP with something else.
As for whether this is good for your protein of interest, smaller peptidic tags are normally better as larger tags may interfere with normal complex/interaction partners.
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