Far western blotting - (Apr/01/2012 )

Hello everyone!

I am trying to work on some protein-protein interactions, and have recently read a few articles and papers that Far western blotting is a very good technique to identify them.
Has anyone used this technique before and could help me describe it for me so that I can get a fair idea on how to do it please?

Thanks
D

-Divya Pothula-

it is similar to immnoblot but w/o non-antibody protein; recombinant protein to be tested may carry a tag to be detected by antibody, or you use an antibody directed against bound protein

-Inmost sun-

see this wikipedia explanation: far western blotting.

-mdfenko-

Thnks all! I am gathering information on it and trying my first trial soon.

-Divya Pothula-

Good luck! I started FWB with radioactively autophosphorylated (purified) kinase; it worked very well but a control for non-phosphorylated form was lacking; so I went ahead with antibodies....

-Inmost sun-

Thanks I have not tried this yet...but then I am very much willing to...as I recently started my PhD...I am firstly trying to purify my recombinant protein and then tag it but not radioactively..some other options... well still working on these.

-Divya Pothula-