2D peptide mapping

[Darren Tyson]

Roney,

I'm planning on doing the identical studies.  A highly recommended
apparatus is Pharmacia's Multiphor II system.  I don't have direct
experience using this equipment, but a close friend swears by it, and
his results look great.

Our lab has BioRad's MiniProtean system for SDS-PAGE and have recently
purchased their insert for tube gel vertical IEF.  If your proteolytic
fragments are >500 Da, they can be separated on 16% Tris/Tricine
SDS-PAGE in the second dimension (after IEF in the first).  This is the
method I intend to use (trypsinizing protein as I write this).

Another alternative is using TLE (electrophoretic TLC) in one dimension
and TLC in the other on common cellulose acetate TLC plates.

For specific protocols you can read Chapters 8 and 23 both by Ralph Judd
in _Methods in Molecular Biology_, Vol 32: Basic Protein and Peptide
Protocols.  Edited by: J. M. Walker.  Copyright 1994.  Humana Press.

Hope this helps,

Darren
-- 
Darren Tyson, Grad Student Cell and Molecular Biology
Saint Louis University, St. Louis, MO
tysondr at slu.edu or darren at primary.net
http://scimart.com/darren/
soon to be moved to
http://www3.primary.net/~darren/