dot plot and overlay - protein-protein interaction (May/26/2011 )

hello,i'm Sarah


In order to study interactions between 2 proteins, i'm going to do a dot plot so i will immobilise my first protein on the nitrocellulose membrane then i will do an overlay , i'm looking for a complete protocol of the overlay and informations about this technique. i found articles about overlay in which they do a an electrophoresis and a transfer on the nitrocellulose membrane. so, i can't do this. could some one help me to find a way to do may dot plot and my overlay. i did not find the concentration of proteins that i have to use.
i hope that someone help me.
thank you very much.

dot blots are performed the same way as westerns except that you apply sample directly to the membrane instead of transferring from a gel.