internal peptide sequencing

[kraepiel]

Some time ago there was a discussion about the best way to go about
digesting a protein to obtain internal peptide sequence. Of course I was
not paying attention then and now I need the information.
	 I have a partially purified protein preparation and can with which I
can get an isolated band of my protein of interest by SDS PAGE.  If I
electroelute this protein, digest it and run a gel to obtain isolated
fragments I will probably lose a lot during these manipulations.  This
protein is pretty low copy number and it is difficult to get enough to
sequnce.  I would be grateful for any alternative strategies one might
use to get internal sequence.


				Todd Lane
				Tlane at princeton.edu