western blot of basic proteins
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
More information about the Methods