Re: Clostripain and Glu-C
Joseph Fernandez (fernaj@rockvax.rockefeller.edu)
Fri, 16 Oct 1998 08:06:54 -0400 (EDT)
Karen and Dave,
We have used both clostripain (Promega) and endoproteinase Glu-C
(Boehringer Mannheim) intermitantly over the past 8 years. We have
observed cleavages at the C-terminal side of ASP as well as GLU using our
standrard hydrogenated triton/tris buffer, but never a trypsin clip.
Clostripain has been used less frequently but has always been dependable to
cleave at ARG, with one exception where we observed a c-terminal lys. We
have used clostripain to get overlapping peptides to compare with LYS-C
digests and have definitely observed internal lysines in these peptides.
Neither enzyme we have had a need to purchase the past year, so
perhaps what you are seeing is a current problem. Wish I had a more
concrete answer, but the enzymes have always worked well for us.
Joe
Joseph Fernandez
Associate Director
The Rockefeller university
Protein/DNA Technology Center
1230 York Ave. New York, NY 10021
Phone: (212)-327-8869
FAX : (212)-327-8620
email: fernaj@rockvax.rockefeller.edu
Lab Web Page: http:\\pdtc.rockefeller.edu