Far western - (Mar/03/2005 )
I'm searching someone who's a master in the technique of Far Western! I would like to test an interaction in vitro between two proteinn but I don't find the good conditions for it! If someone would have a good protocole? Thanks a lot
maybe may you have a look at thos web page :
http://www.utoronto.ca/krause/Farwestern.html (used in scwartz and al. EMBOJ and guichet and al. Nat)
adapted at this webpage :
hope i've been helpful for a few.
you are searching for help with your far-western protocol, yes? Have you thought of using other systems to check for protein-protein interaction like yeast-two-hybrid, (co)-immunoprecipitation or (GST-) pulldowns?
if you could be a little more specific what your problem is, we might be able to give you some advice, maybe!
and please try to avoid double posting, though posting the same topic in two diffent languages is quite innovative!
For the double posting, I realized I didn't write in english but too late...
For the Far Western: The interaction I have to test has been shown in vivo by double hybrid system. I tried a co-IP system last year but no result with interaction (I have shown another one but not this interaction!) We think that Far Western would be a good argument because I use proteins expressed in human cells (cells extracts) and translated in vitro (better than the protein expressed in Coli and more or less purified).
Hope this informations will tell you something!