protein interaction

[houlinlin]

HELP!
Does anyone know the methods which can detect the interaction between proteins the most accurately?

[mith]

Yeast 2 hybrid?

[s3u6e1]

CO-IP?

[MrMistery]

haven't done either of them, but i think co-immunoprecipitation is easier to do. Less places where you can screw up the protocol.

[s3u6e1]

I have tried the 2-hybrid before, it was not too complicated in fact. but at last I didn't get the result I was expecting for. maybe it was because my proteins are secreted protein, we know for 2-hybrid, the reporter genes can only be motivated when the two proteins of interest banding to the AD and BD separately enter into the nuclear. so maybe the 2-hybrid has some limits I think.

For the Co-IP, we need anti-bodies for the proteins we want to test, which may take some time. in order to avoid producing the anti-bodies, I chose to tag my proteins with different tags (3HA,GST,GFP,Myc...), and so I can use the antibody anti-tags which can be purchased from the bio-companies. but now I have a big problem with taging my protein, I am also looking for the solutions now.

I have a paper concerning of the Co-IP, with a detail protocol.

Far3 and Five Interacting Proteins Prevent Premature Recovery from Pheromone Arrest in the Budding Yeast Saccharomyces cerevisiae. Hilary A. Kemp and George F. Sprague, Jr. MOLECULAR AND CELLULAR BIOLOGY, Mar. 2003, p. 1750–1763

I hope that there are something you needs.