western blot of basic proteins

Carl Mayers cnm at mole.bio.cam.ac.uk
Sat Aug 22 07:19:01 EST 1998


I've had similar problems trying to blot basic proteins to 
nitrocellulose.  What worked for me was switching to a more basic 
blotting buffer (3mM Na2Co3, 10mM NaHCo3, 20% (v/v) Ethanol, see Dunn, 
S.D. (1986) Analytical Biochem., 157, 144-153).  I do my blots in this 
buffer in Biorad Mini-Trans blot apparatus, for 2h at 80V.  The buffer 
can get quite hot, so I stand the setup in bucket of ice.
My protein has a predicted pI of around 10, and this works fine for me.
Hope this helps.
All the best, Carl





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