2D peptide mapping
tysondr at sluvca.slu.edu
Mon Feb 17 13:41:06 EST 1997
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 Tyson, Grad Student Cell and Molecular Biology
Saint Louis University, St. Louis, MO
tysondr at slu.edu or darren at primary.net
soon to be moved to
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