n terminal deblocking of protein in gel or pvdf membrane

[Jim Cheetham]

Hello
>     
>     sometimes I have to deal with proteins electroblotted onto pvdf 
>     membrane or in sds page that are impossible to sequence because they 
>     are blocked at the N terminal end .
>     I usually try CNBr cleavage but after that I have problems in the 
>     extraction of obtained peptides.

The protein sequencing lab here routinely sequences from electroblotted
proteins.
You can look up:  Fernandez, J., et al. (1992) Analytical Biochemistry, 201,
255-264.

There is also a more recent reference but it was buried under a desk avalanche
a few days ago, and I cannot find it right now (also by Joe Fernandez, et al.).

Good luck,

Jim

James J. Cheetham
Laboratory of Molecular
and Cellular Neuroscience
The Rockefeller University
cheethj at rockvax.rockefeller.edu