Should we be able to get tryptic digestion (using, say Ulf Hellman's
protocol) and retrieve fragments from proteins bands in this gel--or do
these stain and destain conditions "fix" the protein in the gel so well
that we will not be able to get either digestion of the protein
by the trypsin or extraction of the fragments from the gel pieces.
We actually are trying to do CNBr cleavage of the proteins in-gel--as we
know the chemical cleavage can occur--but are not having good luck
finding fragments by MALDI. CNBr fragmentation of protein in solution
worked well with good MALDI results.
In the coomassie blue stained gel, we are having trouble removing the
coomassie blue--is that a result of our staining method?
Thanks,
Deb McMillen
Institute of MOlecular Biology
University of Oregon
Eugene OR