Investigating protein-protein interactions by far-Westerns.
The identification of protein interaction partners can often elucidate the function of the protein under investigation based on the "guilty by association" concept. Furthermore, the binding event between two proteins can be used as a functional assay when no such assay is available. Despite the large number of advanced techniques that are currently available for studying protein-protein interactions, far-Westerns or blot overlays are still very commonly used in the average laboratory setting due to their powerfulness. This is due to the simplicity and clarity in the results that they produce. Here, the details and mechanics of far-Westerns are discussed to help the reader choose amongst the different variations that exist depending on the question being investigated and the materials available to them. Some examples involving unique questions are also discussed in order to educate the reader on the versatility of far-Westerns. Finally, a troubleshooting section provides the reader with an understanding of the common problems that can be encountered and how these problems can be circumvented.
Chan
Catherine S
CS
Department of Biological Sciences, University of Calgary, 2500 University Dr. NW, T2N 1N4, Calgary, AB, Canada.
Winstone
Tara M L
TM
Turner
Raymond J
RJ
D015153:Blotting, Western / Q000295:instrumentation* / Q000379:methods* / Q000639:trends; D025941:Protein Interaction Mapping / Q000295:instrumentation* / Q000379:methods* / Q000639:trends